Kev Sib Piv Ntawm Ob Qhov Kev Tshawb Fawb Txog Kev Tshawb Fawb Txog Kev Tshawb Fawb Ntawm Cov Ntshav Qab Zib Cov Tshuaj Tiv Thaiv Kab Mob rau Porcine Kab Mob Kab Mob Kab Mob
Oct 11, 2023
Abstract:
Kev tiv thaiv Lactogenic yog qhov tseem ceeb rau kev tiv thaiv piglets tiv thaiv ntau yam kab mob nrog rau cov kab mob porcine kab mob raws plab. Circulating neutralizing antibody qib hauv sow sera tuaj yeem pab txiav txim siab seb puas muaj cov tshuaj tiv thaiv kab mob tiv thaiv kab mob tuaj yeem tiv thaiv piglets. Neutralizing antibodies tuaj yeem kuaj pom los ntawm ntau yam kev kuaj mob. Txoj kev tshawb no tau soj ntsuam cov yam ntxwv ntawm kev kuaj mob ntawm ob qhov kev tshuaj ntsuam xyuas cov tshuaj tiv thaiv kab mob rau cov kab mob porcine kab mob raws plab kab mob-neutralizing cov tshuaj tiv thaiv hauv cov ntshav ntawm cov gilts sib tw. Plaub pawg kho mob, kev tswj hwm, tsis muaj tshuaj tiv thaiv, txhaj tshuaj tiv thaiv ua ntej kev sib tw, thiab txhaj tshuaj tiv thaiv tom qab kev sib tw, muaj 20 gilts. Cov qauv kuaj ntshav tau sau los ntawm txhua qhov gilt ua ntej thiab tom qab qhov kev sib tw nrog tus kab mob porcine kab mob raws plab. Cov qauv raug soj ntsuam rau qhov muaj cov tshuaj tiv thaiv kab mob nruab nrab los ntawm kev tshuaj ntsuam fluorescent tsom mus rau nruab nrab thiab kev ntsuam xyuas siab dhau los ntawm qhov nruab nrab. Diagnostic rhiab heev thiab tshwj xeeb rau fluorescent tsom xam neutralization thiab high-throughput neutralization assays rau txoj kev tshawb no tau optimized ntawm ib tug txiav tawm ntawm ib tug dilution ntawm 80 thiab 80% fluorescent txo raws li thiab pom tau hais tias muaj kev pom zoo raws li kappa statistic. Kev tsom xam fluorescent neutralization thiab cov kev ntsuam xyuas siab dhau los ntawm qhov nruab nrab tuaj yeem siv los saib xyuas cov xwm txheej ntawm cov tshuaj tiv thaiv kab mob hauv cov tsiaj los yog cov pej xeem ntawm cov tsiaj. Qhov kev ntsuam xyuas siab dhau los muaj qhov zoo tshaj qhov kev ntsuam xyuas fluorescent hauv qhov uas nws muaj qhov tshwj xeeb siab dua ntawm qhov kev txiav tawm thiab qhov xwm txheej ntawm cov txiaj ntsig tso cai rau kev ntxub ntxaug ntau dua ntawm tus neeg tau txais txiaj ntsig.
cistanche ntxiv cov txiaj ntsig-nce kev tiv thaiv
Keywords: PEDV; neutralizing antibody; kev kuaj mob
1. Taw qhia
Porcine epidemic diarrhea virus (PEDV) yog Alphacoronavirus uas tau dhau los ua neeg nyob hauv Asmeskas tom qab nws tshwm sim hauv Tebchaws Meskas xyoo 2013 [1–3]. Biosecurity ntsuas yog tsom rau kev khaws cov kab mob xws li PEDV tawm ntawm swine herds kom txo tau qhov cuam tshuam ntawm tus kab mob [4]. Kev ntsuam xyuas Serology yog ib qho cuab yeej tseem ceeb los txiav txim seb cov npua yav dhau los tau raug thiab kis los ntawm cov kab mob. Qhov tshwm sim tuaj yeem tshwm sim ib txwm muaj (piv txwv li, noj cov khoom noj uas muaj tus kab mob) lossis txhob txwm (xws li, koom nrog cov tsiaj txhu hauv cov tsiaj, qhov ncauj qhov ntswg, lossis plab hnyuv). Rau lub hom phiaj ntawm txoj kev tshawb no, kev nthuav tawm tau ua tiav los ntawm kev sib tw nrog PEDV-zoo cov ntaub so ntswg homogenate los ntawm kev pom zoo, kev kuaj mob ntawm PEDV. Nkauj et al. [5] pom tau tias cov tshuaj tiv thaiv kab mob tsis sib haum xeeb tsis txawv ntawm cov noob qoob loo tau muab tshuaj tiv thaiv qhov ncauj lossis intramuscular Vero cell attenuated PEDV DR13 tshuaj tiv thaiv. Txoj kev tshawb no kuj pom tau hais tias cov tshuaj tiv thaiv qhov ncauj qhov ncauj tau txo qis kev tuag ntawm piglet piv rau kev txhaj tshuaj intramuscular. Tsis tas li ntawd, de Arriba et al. [6] tau qhia txog kev sib raug zoo ntawm IgA thiab IgG antibody-secreting cells (ASC) nyob rau hauv cov ntshav thiab plab-txo cov ntaub so ntswg lymphoid. Lub diffuse, epitheliochorial placenta ntawm sows tiv thaiv lub cev tiv thaiv kab mob thiab cov tshuaj tiv thaiv los ntawm kev xa mus rau piglets hauv utero ua npua agammaglobulinemic thaum yug los [7]. Kev tiv thaiv niam txiv yog kis los ntawm cov noob rau lawv cov piglets ntawm mammary secretions thiab ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv cov kab mob enteric xws li PEDV, TGEV, thiab rotavirus. Yog li ntawd, piglets cia siab rau colostrum thiab mis nyuj cov tshuaj tiv thaiv rau kev tiv thaiv los ntawm cov kab mob no. Serum immunoglobulins yog ib tug loj contributor rau colostrum whereas immunoglobulins nyob rau hauv cov mis nyuj yog tsim nyob rau hauv lub mammary caj pas [8]. Yog li, kev ntsuam xyuas cov tshuaj tiv thaiv kab mob hauv nruab nrab tuaj yeem pab txiav txim siab seb puas muaj lub cev tiv thaiv kab mob hauv lub cev ua ntej sow sera tuaj yeem muab kev tiv thaiv rau piglets. Cov txheej txheem ntawm neutralizing cov tshuaj tiv thaiv muaj xws li kev sib sau ua ke, inhibition ntawm tus kab mob nkag mus rau ntawm lub hom phiaj ntawm tes, thiab inhibition ntawm tus kab mob replication nyob rau hauv lub hom phiaj cell [9]. Kev tshuaj ntsuam xyuas tau tsim los txhawm rau txheeb xyuas cov tshuaj tiv thaiv tsis zoo rau PEDV suav nrog tus kab mob nruab nrab (VN), fluorescent focus neutralization (FFN), thiab high-throughput neutralization test (HTNT) [2,10–13]. Raws li tau piav qhia los ntawm Sarmento et al. [2], FFN assay yog ib qho kev txhim kho raws li VN assay raws li qhov kev ntsuam xyuas yog rhiab heev vim hais tias qhov kev ntsuam xyuas yog raws li kev tshawb pom ntawm tus kab mob antigens qhia nyob rau saum npoo ntawm cov kab mob Vero hlwb thaum lub VN assay yog raws li cytopathic nyhuv tshwm sim. los ntawm tus kab mob. Tsis tas li ntawd, FFN cov txiaj ntsig tuaj yeem tau txais sai dua li VN cov txiaj ntsig vim tias cov nyhuv cytopathic yuav siv sij hawm ntev dua los soj ntsuam piv rau cov kab mob txuas nrog FFN. Qhov kev ntsuam xyuas HTNT yog ib qho kev txhim kho ntxiv ntawm qhov kev ntsuam xyuas FFN raws li qhov kev ntsuam xyuas HTNT tso siab rau cov duab digital cytometry es tsis yog kev txhais cov kws tshaj lij kom tau txais cov txiaj ntsig. Yog li, tso cai rau tag nrho cov phaj nyeem hauv tsawg dua 3 min, ua kom muaj kev ntsuas ntau hauv fluorescent siv, thiab muaj peev xwm khaws cov duab rau kev tshuaj xyuas. Yog li, HTNT zoo nkaus li tsim nyog siv rau hauv kev lag luam swine herds. Txoj kev tshawb fawb tam sim no tau muab piv rau cov yam ntxwv ntawm kev kuaj mob (rhiab heev thiab qhov tshwj xeeb) thiab qhov zoo thiab qhov tsis zoo ntawm qhov kev kwv yees, ntawm ob qhov kev ntsuam xyuas cov tshuaj tiv thaiv kab mob rau PEDV, FFN, thiab HTNT. Qhov no tsim los ntawm kev ua haujlwm ntawm Sarmento li al. [2] los ntawm kev sim cov neeg tsis muaj zog, muaj kab mob, thiab txhaj tshuaj tiv thaiv tsiaj txhawm rau pab txheeb xyuas qhov kev ntsuam xyuas siv rau hauv qhov chaw kuaj mob siab tshaj plaws. Raws li cov txiaj ntsig ntawm Sarmento et al. [2], peb qhov kev xav tsis zoo yog tias tsis muaj qhov sib txawv ntawm qhov kev kuaj mob ntawm ob qhov kev ntsuam xyuas kom pom cov tshuaj tiv thaiv PEDV hauv cov ntshav ntawm PEDV-kev sib tw gilts.
cistanche ntxiv cov txiaj ntsig-nce kev tiv thaiv
2. Cov ntaub ntawv thiab cov txheej txheem
2.1. Qhov chaw thiab xaiv Swine
Raws li yav dhau los tau piav qhia los ntawm Brown li al. [14], txoj kev tshawb no tau siv 4 pawg kho mob hu ua Kev Tswj, NV (tsis txhaj tshuaj tiv thaiv), Pre ( txhaj tshuaj tiv thaiv ua ntej kev sib tw), thiab Post ( txhaj tshuaj tiv thaiv tom qab kev sib tw). Txhua pawg tau pib tsim los ntawm 20 kev lag luam, crossbred, gilts. Tag nrho ntawm 60 gilts tau yooj yim xaiv los ntawm kev lag luam swine ua liaj ua teb hauv nruab nrab Iowa uas tsis muaj keeb kwm ntawm PEDV. Lub 20 gilts hauv Pawg Tswjhwm tau rov qab siv rau pawg NV. Pab pawg NV tsuas yog muaj 18 tus npua xwb vim ob lub gilts raug tshem tawm ntawm txoj kev tshawb no vim muaj teeb meem tsis cuam tshuam nrog PEDV kev sib tw.
Raws li cov pov thawj ntxiv ntawm PEDV qhov xwm txheej tsis zoo, 12 mL ntawm cov ntshav tau sau los ntawm jugular venipuncture siv lub ntsuas 16- ntsuas, 1.5- ntiv tes koob thiab 12 mL koob txhaj tshuaj. Cov ntshav tau sau los ntawm cov ntshav kuaj los ntawm centrifugation ntawm 1.500 × g rau 8 min (min), muab faib ua 1 mL aliquots, thiab khaws cia ntawm −80 ◦C. Cov qauv ntshav tau kuaj nrog wv-ELISA ntawm Iowa State University Veterinary Diagnostic Laboratory (ISU-VDL, Ames, IA, USA) [15].
2.2. Kev Muab Tsiaj Rau Cov Pab Pawg Kho Mob
Lub rau caum gilts tau muab faib ua 3 pawg, txhua tus muaj 20 tus neeg lag luam, cov poj niam sib tw. Txhawm rau tsim plaub pawg kho mob, 20 gilts tau siv ua ntu zus rau ob pawg kho mob, CONTROL thiab NV. Rov qab siv 20 gilts tso cai rau kev txhim kho thiab tswj cov pab pawg ntawm PEDV-zoo gilts uas tau raug rau PEDV thiab txo tag nrho cov tsiaj txhu siv hauv txoj kev tshawb no. Qhov seem 40 gilts suav nrog PRE thiab POST pawg, nrog 20 gilts hauv txhua pab pawg. Lub sijhawm ntawm cov xwm txheej los ntawm pab pawg kho mob tau piav qhia hauv Brown li al. [14]. Hauv luv luv, ib tus neeg kuaj ntshav tau sau ib zaug hauv ib lub lim tiam hauv txhua pab pawg kho mob tom qab qhov kev sib tw. Kev piav qhia ntxiv yog tham hauv Tshooj 2.4 thiab 2.5. Pab pawg kho mob "PRE" gilts tau txhaj tshuaj tiv thaiv ua ntej tuaj txog ntawm qhov chaw sab nraud. Pab pawg kho mob "POST" gilts tsis tau txhaj tshuaj kom txog thaum tom qab kev sib tw.
2.3. Tsev thiab Kev Saib Xyuas Swine
Thaum tuaj txog ntawm qhov chaw tshawb fawb, gilts tau nyob hauv pab pawg kho mob: tag nrho 20 gilts hauv pab pawg kho mob tau nyob hauv ib tus cwj mem nrog ad libitum pub thiab dej. Tag nrho cov cwj mem nyob hauv tib chav. Ib tus cwj mem raug tso tseg ntawm NV PRE thiab POST pawg kom txo qis kev sib kis. Pub, dej, chav tsev kub siab thiab qis, thiab cov av noo siab thiab qis tau raug saib xyuas kom ntseeg tau tias muaj kev sib raug zoo nrog cov qauv tsim khoom. Kev soj ntsuam kev noj qab haus huv tau ua thiab sau tseg ib hnub ib zaug los soj ntsuam kev mob plab, qaug zog, mob plab, thiab lwm yam mob tshwm sim. Ib pob pob ntseg (Integra Hog, Allflex, Dallas, TX, USA) tau muab tso rau hauv pob ntseg sab xis ntawm txhua qhov gilt rau kev txheeb xyuas tus tsiaj.
2.4. Qhov ncauj sib tw
Tom qab lub sijhawm 4-hnub acclimation, txhua qhov gilt hauv CONTROL tau pom qhov ncauj rau PEDV siv cov ntaub so ntswg homogenate sau los ntawm kev kuaj pom muaj tus kab mob PEDV ntawm kev lag luam ua liaj ua teb. Cov homogenate tau ua raws los ntawm Iowa State University Veterinary Diagnostic Laboratory thiab pom tias yog US tus qauv qauv, genogroup G2b. Kaum milliliters ntawm homogenate tau tov nrog 590 mL ntawm phosphate buffered saline thiab 30 mL ntawm qhov sib tov tau muab tshuaj ironically rau txhua lub gilt. Lub inoculum tau lees paub PEDV zoo los ntawm lub sijhawm tiag tiag, rov qab transcriptase polymerase saw cov tshuaj tiv thaiv nrog CT tus nqi ntawm 19.6 thiab 14.23 × 107 genomic luam ib mL thiab kev loj hlob raws plab hauv gilts tom qab kev sib tw. Cov neeg kuaj ntshav tau sau txhua 7 hnub tom qab qhov kev sib tw ntawm jugular venipuncture raws li tau piav qhia saum toj no mus txog hnub 42 tom qab kev sib tw. Cov ntshav tau raug sau thiab khaws cia raws li tau piav qhia saum toj no thiab cov qauv raug xa mus rau ISU VDL txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau nws cov kab mob Hnub 60 tom qab pib qhov kev sib tw ntawm CONTROL, qhov seem 40 gilts tau coj mus rau qhov chaw tshawb fawb thiab 18 ntawm CONTROL gilts tau dhau los ua NV. Ob lub gilts raug tshem tawm ntawm CONTROL vim yog cov xwm txheej tsis cuam tshuam nrog kev kawm. Tom qab lub sijhawm 3-hnub tom qab tuaj txog, 18 gilts tau rov sib tw (NV) thiab 40 gilts (PRE thiab POST) tau sib tw, kuaj ntshav tau sau, cov ntshav tau sau, thiab cov qauv xa mus kuaj raws li tau piav qhia hauv qab no .
2.5. Kev txhaj tshuaj ntawm Gilts
Plaub caug gilts tau nyob ntawm thaj chaw ntawm keeb kwm thiab raug coj mus rau qhov chaw tshawb fawb 8.5 lub lis piam tom qab tuaj txog ntawm CONTROL. Nees nkaum gilts tau txhaj tshuaj tiv thaiv 5 thiab 2 lub lis piam ua ntej qhov kev sib tw, raws li kev pom zoo los ntawm lub tuam txhab tsim cov tshuaj tiv thaiv, nrog rau kev muag khoom, tua cov tshuaj tiv thaiv PEDV, genogroup G2b, (Porcine Epidemic Diarrhea Vaccine, Zoetis, Inc., Florham Park, NJ , USA) suav nrog PRE pawg. Qhov seem nees nkaum tau txhaj tshuaj tiv thaiv tib yam, 1 thiab 3 lub lis piam tom qab kev sib tw PEDV suav nrog POST pawg. Kaum yim ntawm thawj nees nkaum CONTROL gilts tseem nyob hauv txoj kev tshawb fawb thiab suav nrog NV pawg thiab tsis tau txais cov tshuaj tiv thaiv rau PEDV
cistanche tsob nroj-nce kev tiv thaiv kab mob
2.6. Neutralizing Antibody Assay
2.6.1. Fluorescent Focus Neutralization (FFN) txheej txheem
FFN tau ua los ntawm Iowa State University Veterinary Diagnostic Laboratory, tus txheej txheem kev ua haujlwm 9.5324 [16]. Luv luv, Vero 76 hlwb tau npaj los ntawm kev yaug thiab incubating nrog Trypsin-Ethylenediaminetetraacetic acid (EDTA) rau 5 feeb ntawm 37 ◦C. A 1:20 dilution ntawm cell thiab cell propagation nruab nrab yog tsim thiab 200 µL ntxiv rau txhua lub qhov dej ntawm 96-zoo phaj thiab incubated rau 48 h. Cov phaj tau muab pov tseg thiab yaug nrog tus kab mob inoculation nruab nrab. Cov qauv ntshav tau ua kom sov hauv chav dej ntawm 56 ◦C rau 30 feeb thiab tom qab ntawd diluted hauv 2-fold serial dilution pib ntawm 1:10 hauv lub phaj. Cov kab mob Tshuag tau diluted rau 100-200 foci tsim units ib 100 µL thiab 100 µL ntxiv rau txhua qhov dej thiab incubated rau 60 min ntawm 37 ◦C nrog 5% carbon dioxide (CO2). Cov phaj xeem tau muab pov tseg, yaug thiab 150 µL ntawm cov kua qaub / kab mob sib xyaw ntxiv rau cov qhov dej sib txuas thiab incubated rau 60 min ntawm 37 ◦C. Cov kua qaub / kab mob sib xyaw tau muab pov tseg thiab cov qhov dej tau ntxuav nrog tus kab mob inoculation nruab nrab. 150 µL ntawm tus kab mob inoculation nruab nrab tau ntxiv rau txhua qhov dej thiab incubated rau 24 teev ntawm 37 ◦C thiab 5% CO2. Media tau raug tshem tawm ntawm lub phaj thiab cov hlwb raug kho los ntawm kev ntxiv 80% acetone, uas tau muab tso rau hauv ib da dej dej khov, rau 15 feeb. Acetone tau muab pov tseg thiab lub phaj yog cua qhuav. A 1:100 dilution ntawm FITC conjugated PEDV NP monoclonal antibody diluted hauv PBS (pH 7.2) tau npaj thiab 50 µL tau ntxiv rau txhua qhov dej thiab incubated ntawm 37 ◦C rau 60 min. Lub phaj tau ntxuav nrog PBS thiab tom qab ntawd nyeem siv lub tshuab ntsuas fluorescence (Olympus BX51, 10X lub hom phiaj, FITC lim). Cov qhov dej zoo tsis pom cov fluorescing plaques thiab tsis pom cov plaques ntsuab. Neutralizing antibody titers tau hais tias yog qhov siab tshaj plaws hauv cov ntshav dilution nrog 90% txo tus kab mob rov ua dua piv rau kev tswj tus kab mob. Cov qauv uas muaj titer Ntau dua lossis sib npaug rau 20 tau muab cais ua qhov zoo rau PEDV-neutralizing antibodies. Lub titer siab dua qhia tias muaj ntau dua ntawm cov tshuaj tiv thaiv kab mob hauv cov qauv. Ib qho piv txwv ntawm cov qauv zoo thiab tsis zoo sera muaj nyob rau hauv cov ntaub ntawv ntxiv.
2.6.2. High Throughput Fluorescent Neutralization Test (HTNT) txheej txheem
HTNT tau ua los ntawm Iowa State University Veterinary Diagnostic Laboratory, tus txheej txheem kev khiav hauj lwm 9.5324 Version 2 [17]. Cov qauv kuaj ntshav tau ua kom sov hauv chav da dej ntawm 56 ◦C rau 30 feeb. Tus kab mob Tshuag tau serial diluted 10-fold hauv kab mob inoculation nruab nrab (DMEM nrog 2 µg/mL Trypsin-TPCK), kis mus rau Vero 81 hlwb (ATTC® CCL-81™), thiab incubated ntawm 37 ◦C nrog 5% CO2 rau 5 hnub. Qhov kawg dilution nyob rau hauv uas cov nyhuv cytopathic tau pom tau siv los txiav txim qhov nruab nrab cov ntaub so ntswg kab lis kev cai kis kab mob (TCID50) siv txoj kev Spearman thiab Karber [18]. Ib puas microliters ntawm cov kua cell (Cell propagation Medium, Trypsin-EDTA, thiab Vero 81 hlwb), yog diluted nrog 100 µL ntawm cell propagation nruab nrab (DMEM nrog 10% FBS thiab 1% penicillin-streptomycin) ua 1: 1 dilution . Lub hemocytometer tau siv los suav thiab suav cov dilution los ua 5 × 104 cell / qhov dej thiab ntxiv rau txhua qhov dej ntawm lub qhov dej ntshiab hauv qab dub 96-zoo phaj (Coring 96 well CellBind microplate, Sigma) thiab incubated rau 48 h ntawm 37 ◦C nrog 5% CO2 los ua kom cov cell sib txuas (100% confluent). Txhua lub phaj muaj kev tswj zoo, kev tswj tsis zoo, thiab kev tswj tus kab mob. Tom qab ntawd, tsib microliters ntawm cov cua sov-inactivated tswj thiab cov qauv tau muab tso rau hauv cov qhov dej sib npaug kom ua 1:40 dilution los ntawm thawj 1:20 los ntawm kev ntxiv 95 µL ntawm tus kab mob inoculation nruab nrab, ces 100 µL ntawm 1:10 diluted PEDV tus kab mob (3.16 × 105. TCID 50 / ml). Cov kua qaub / cov kab mob sib xyaw yog incubated rau 1 h ntawm 37 ◦C nrog 5% CO2. Tag nrho cov cell qhov dej ntawm lub phaj kuaj tau ntxuav 3 zaug nrog 150 µL ntawm cell ntxuav nruab nrab (DMEM nrog 1% penicillin-streptomycin), thiab 150 µL ntawm cov tshuaj tiv thaiv kab mob / cov kab mob sib xyaw thiab cov kab mob tswj tau raug xa mus rau lub phaj kuaj thiab incubated rau 90 min. ntawm 37 ◦C. Cov kua qaub / cov kab mob sib xyaw tau muab pov tseg thiab cov phaj tau ntxuav ib zaug nrog lub tshuab ntxuav tes, raws li tau piav qhia saum toj no, ua raws li ib zaug nrog cov kab mob inoculation nruab nrab. Tus kab mob inoculation nruab nrab (150 µL) tau ntxiv rau txhua qhov dej thiab incubated rau 24 h ntawm 37 ◦C nrog 5% CO2. Tus kab mob inoculation nruab nrab tau muab pov tseg thiab lub phaj tau ntxuav nrog PBS pH 7.4. Cells raug kho los ntawm kev ntxiv 4 ◦C, 80% acetone rau txhua qhov dej thiab incubated ntawm chav tsev kub rau 15 min. Tom qab ntawd Acetone raug tshem tawm thiab lub phaj yog cua qhuav rau 30 feeb. Txhua qhov dej tau yaug nrog PBS pH 7.4. 50 µL ntawm FITC-conjugated PEDV NP monoclonal antibody (SD6-29 clone, Medgene Labs) diluted 1:100 hauv PBS pH 7.4 ntxiv rau txhua qhov dej thiab incubated rau 60 min ntawm 37 ◦C. Lub phaj tau ntxuav plaub zaug nrog PBS pH 7.4, tawm hauv lub phaj kawg rau 5 feeb ces muab pov tseg thiab hloov nrog 100 µL ntawm PBS pH 7.4. Cov phaj tau nyeem nrog SpectraMax i3 × siv SoftMax Pro 6.5 (Molecular Devices, San Jose, CA, USA) siv 30 ms raug lub sij hawm thiab 20 um focal hloov kho rau 541 wavelengths. Cov qauv nrog Tag Nrho Fluorescence Reduction (% FR) Ntau dua lossis sib npaug li 85% tau muab cais ua qhov zoo rau cov tshuaj tiv thaiv tsis zoo [2]. Tag nrho cov fluorescence txo tau raug xam raws li 100 − ([Nruab nrab piv txwv Total Fluorescence Intensity/Average Negative Control Total Fluorescence Intensity]) × 100). Kev txo qis tag nrho cov fluorescence ntau dua qhia tias muaj ntau dua neutralizing antibody concentrations hauv cov qauv. Ib qho piv txwv ntawm cov qauv zoo thiab tsis zoo sera muaj nyob rau hauv cov ntaub ntawv ntxiv.
2.7. Diagnostic Characteristics: Sensitivity thiab Specificity thiab Positive and Negative Predictive Values
Diagnostic rhiab heev thiab tshwj xeeb raug xam rau HTNT siv ib % FR txiav tawm ntawm 70, 75, 80, 85, thiab 90 thiab FFN siv cov qauv dilution cutoffs ntawm 20, 40, 80, thiab 160 los txiav txim qhov optimized cutoff. Optimized tau txhais tias yog qhov kev txiav tawm qhov kev kuaj mob tshwj xeeb yog ua kom tiav nrog qhov tsawg kawg ntawm kev kuaj mob tshwj xeeb. Ib pawg ntawm xya caum-rau qhov paub PEDV-tsis zoo qauv raug xaiv los ntawm thawj ob lub ntsiab lus piv txwv rau CONTROL thiab POST pawg. 36 paub PEDV-zoo qauv raug xaiv los ntawm ob lub ntsiab lus piv txwv kawg rau pawg NV. Ib qho kev txiav tawm ntawm ntau dua 85 tau siv los txheeb xyuas cov qauv zoo, ua ke nrog ISU VDL tus qauv kev ua haujlwm rau kev ntsuas [17]. Diagnostic rhiab heev tau kwv yees los ntawm kev faib cov naj npawb ntawm cov qauv paub zoo uas kuaj pom zoo los ntawm tag nrho cov qauv paub zoo. Kev kuaj mob tshwj xeeb tau kwv yees los ntawm kev faib cov lej paub tsis zoo uas kuaj pom qhov tsis zoo los ntawm cov lej paub tsis zoo. Qhov kev txiav tawm uas ua kom zoo tshaj qhov kev kuaj mob tshwj xeeb thiab qhov rhiab heev raug xaiv rau kev sib piv. Rau lub hom phiaj ntawm txoj kev tshawb no, diagnostic specificity (DSp) thiab rhiab heev (DSe) tau maximized, qhov tseeb tus nqi ntawm qhov sib txawv ntawm rhiab heev thiab tshwj xeeb qhov tseem ceeb yog tsawg tshaj plaws. Tus nqi kwv yees zoo (PPV) tau kwv yees los ntawm kev faib cov lej ntawm cov qauv paub zoo uas kuaj pom qhov zoo (TP) los ntawm tag nrho cov qauv kuaj pom zoo (TP ntxiv rau qhov tsis zoo [FP]). Kev kwv yees tus nqi tsis zoo (NPV) tau kwv yees los ntawm kev faib cov lej paub tsis zoo (TN) uas kuaj tsis zoo los ntawm tag nrho cov qauv kuaj tsis zoo (TN ntxiv rau qhov tsis zoo tsis zoo [FN]).
cistanche tubulosa- txhim kho lub cev tiv thaiv kab mob
Nyem qhov no mus saib Cistanche Enhance Immunity khoom
【Nug ntxiv】 Email: cindy.xue@wecistanche.com / Whats App: 0086 18599088692 / Wechat: 18599088692
2.8. Kev txheeb cais
Tsis muaj cov txiaj ntsig tsis paub meej lossis cov ntaub ntawv ploj lawm hauv txoj kev tshawb no. Tag nrho cov ntaub ntawv tau txheeb xyuas siv SAS Version 9.4 (SAS Instit. Cary NC). 609 kev soj ntsuam los ntawm 6{{10}} npua uas yog ib qho ntawm 4 pawg tau siv. Ob qhov txiaj ntsig tau txiav tawm (40 thiab 80) rau FFN qhov kev xeem tau raug txiav txim siab thiab muab piv rau qhov txiav tawm ntawm 85 rau HTNT. Rau txhua tus, ib qho qauv sib xyaw ua ke tau haum los piav qhia cov txiaj ntsig ntawm pab pawg ntawm kev pom zoo ntawm FFN thiab HTNT kev soj ntsuam, nrog rau cov txiaj ntsig tshwj xeeb ntawm npua. Txhua qhov kev sib piv sib piv ntawm cov pab pawg tau ua tiav. Ntxiv mus, rau ob ntawm peb qhov kev txiav tawm qhov tseem ceeb siv rau FFN kev soj ntsuam, Cohen's Kappa coefficient raug xam los txiav txim siab tag nrho cov kev pom zoo ntawm FFN thiab HTNT kev soj ntsuam. Kappa coefficients tau txhais raws li hauv qab no: tsawg dua 0.0, "poor" pom zoo; nruab nrab ntawm 0.0 thiab 0.20, "me ntsis" kev pom zoo; nruab nrab ntawm 0.21 thiab {{30}}.40, "fair" kev pom zoo; nruab nrab ntawm 0.41 thiab 0.60, "nruab nrab" kev pom zoo; nruab nrab ntawm 0.61 thiab 0.80, "kev pom zoo"; thiab nruab nrab ntawm 0.81 thiab 1.0, "yuav luag zoo tag nrho" kev pom zoo [19]. Qhov tseem ceeb tau teeb tsa ntawm p Tsawg dua lossis sib npaug rau 0.05. Lub thawv thiab whisker plots tau tsim los siv R software.
3. Cov txiaj ntsig
Table 1 qhia txog qhov kev kuaj mob rhiab heev thiab qhov tshwj xeeb rau FFN thiab HTNT kev soj ntsuam ntawm qhov kev txiav tawm xaiv. Diagnostic rhiab heev thiab tshwj xeeb rau ob qhov kev ntsuam xyuas rau txoj kev tshawb no tau optimized ntawm ib tug txiav tawm ntawm ib tug dilution ntawm 80 thiab 80% FR feem.
Table 1. Diagnostic rhiab heev (DSe), Diagnostic specificity (DSp), Positive Predictive Value (PPV), thiab Negative Predictive Value (NPV) rau HTNT thiab FFN kev soj ntsuam siv ntau yam kev txiav tawm uas teev nyob rau hauv kab thib ob ntawm lub rooj. Piv txwv li, rau qhov kev ntsuam xyuas HTNT ntawm kev txiav tawm ntawm 70, DSe=0.97 (97%), DSP=0.91 (91%), PPV=0.83 ( 83%), NPV=0.95 (95%). U=tsis tau txhais.
Daim duab 1 qhia txog lub thawv thiab cov ntawv nplawm ntawm HTNT cov txiaj ntsig los ntawm hnub tom qab kev sib tw los ntawm pab pawg kho mob. Cov qauv los ntawm CONTROL thiab POST pawg (n=76) tau muab faib ua qhov tsis zoo nyob rau hnub −4, 0, thiab 7 raws li xav tau vim yav dhau los ntawm cov npua. Pab pawg NV tau tsim cov lus teb anamnestic tom qab qhov kev sib tw. Kev txo qis qhov sib txawv tau tshwm sim ntawm HTNT cov txiaj ntsig raws li pom los ntawm kev txo qis ntawm cov txiaj ntsig hauv ob pawg tshuaj tiv thaiv (PRE thiab POST) tom qab kev sib tw nrog PEDV. Qhov kev poob qis no tuaj yeem pom nyob rau hauv daim duab 1 vim tias cov pob tw thiab cov ntawv tawm ntawm POST thiab PRE-ploj ua luv dua 14 hnub tom qab kev sib tw. Daim duab 2 qhia txog lub thawv thiab cov ntawv nplawm ntawm lub ntuj hloov pauv FFN cov txiaj ntsig los ntawm hnub tom qab kev sib tw los ntawm pab pawg kho mob, uas muaj txiaj ntsig zoo ib yam li HTNT kev soj ntsuam. Ob qhov HTNT (Daim duab 1) thiab FFN (Daim duab 2) cov txiaj ntsig tau qhia cov lus teb rau kev sib tw thiab txhaj tshuaj tiv thaiv raws li pom nrog nce % FR thiab titers los ntawm ob qho kev ntsuam xyuas. Thaum zoo sib xws hauv qhov loj, qhov sib txawv ntawm txhua qhov kev ntsuam xyuas tau raug txo qis tom qab kev sib tw homologous (NV). Txo qhov kev hloov pauv kuj tshwm sim tom qab txhaj tshuaj tiv thaiv (PRE thiab POST). Qhov kev hloov pauv ntawm qhov tsis zoo ntawm cov tshuaj tiv thaiv kab mob los ntawm lub lim tiam tom qab raug txo qis tom qab txhaj tshuaj tiv thaiv thiab kev sib tw homologous nrog PEDV (Figures 1 thiab 2). Muaj qhov zoo sib xws hauv kev txo qis ntawm kev hloov pauv ntawm cov tshuaj tiv thaiv kab mob nruab nrab pom nyob rau hauv ob qho kev ntsuam xyuas cov txiaj ntsig ntawm cov tshuaj tiv thaiv thiab homologous sib tw cov pab pawg kho mob. Kev sib tw homologous ntawm PEDV thiab kev txhaj tshuaj thib ob tau tsim cov lus teb anamnestic uas ua rau muaj qib siab dua ntawm cov tshuaj tiv thaiv kab mob nrog txo qis kev sib txawv ntawm cov npua piv rau qhov kev sib tw ua ntej thiab tom qab txhaj tshuaj tiv thaiv raws li ntsuas los ntawm ob qho kev ntsuam xyuas (Daim duab 1). Raws li kev soj ntsuam ntawm qhov kev kuaj mob rhiab heev thiab qhov tshwj xeeb, qhov kev txiav tawm ntawm 80 (titer) thiab 85 (% FR) raug xaiv los txheeb xyuas qhov kev pom zoo rau FFN thiab HTNT kev soj ntsuam raws. Qhov kev xeem kappa pom tau tias "nruab nrab" kev pom zoo ntawm 0.5257 nrog rau 95% kev ntseeg siab ntawm 0.461, 0.5904 ntawm qhov kev ntsuam xyuas rau kev txiav tawm 80 (titer) thiab 85 (% FR) rau FFN thiab HTNT kev xeem.
Daim duab 1. Box-and-whisker plots ntawm HTNT antibody teb curves los ntawm hnub tom qab kev sib tw rau txhua pab pawg kho mob. Qhov nruab nrab bar sawv cev tus nqi nruab nrab ntawm cov txiaj ntsig los ntawm lub lim tiam. Cov pob khawm (sab saum toj thiab hauv qab ntawm lub thawv) sawv cev rau thawj (Q1) thiab thib peb (Q3) quartiles. Qhov chaw ntxoov ntxoo sawv cev rau qhov sib txawv ntawm qhov sib txawv (IQR). Cov kua txob tau txiav txim siab raws li hauv qab no: Lub ntsej muag sab saum toj=Q3 + 1.5∗IQR; Hauv qab whisker=Q1 + 1.5∗IQR. Yog tias tsis muaj cov ntaub ntawv taw qhia nyob ntawm tus nqi suav, cov ntaub ntawv tom ntej no ze tshaj rau Q1 lossis Q3 yog qhov txwv ntawm lub whisker. Qhib lub voj voog (◦) qhia qhov tsis sib xws. Ib qho kev txiav ntawm 85 yog qhia los ntawm kab khiav ntawm 85 ntawm y-axis.
Daim duab 2. Box-and-whisker plots ntawm FFN antibody teb curves los ntawm hnub tom qab kev sib tw rau txhua pab pawg kho mob. Qhov nruab nrab bar sawv cev tus nqi nruab nrab ntawm cov txiaj ntsig los ntawm lub lim tiam. Cov pob khawm (sab saum toj thiab hauv qab ntawm lub thawv) sawv cev rau thawj (Q1) thiab thib peb (Q3) quartiles. Qhov chaw ntxoov ntxoo sawv cev rau qhov sib txawv ntawm qhov sib txawv (IQR). Cov kua txob tau txiav txim siab raws li hauv qab no: Lub ntsej muag sab saum toj=Q3 + 1.5∗IQR; Hauv qab whisker=Q1 + 1.5∗IQR. Yog tias tsis muaj cov ntaub ntawv taw qhia nyob ntawm tus nqi suav, cov ntaub ntawv tom ntej no ze tshaj rau Q1 lossis Q3 yog qhov txwv ntawm lub whisker. Qhib lub voj voog (◦) qhia qhov tsis sib xws. Ib qho kev txiav ntawm 40 yog qhia los ntawm kab khiav ntawm ln(40) ntawm y-axis.
4. Kev sib tham
Peb xav tias qhov kev hloov pauv txo qis thiab cov lus teb anamnestic cuam tshuam nrog homogenous herd tiv thaiv kab mob thiab kev tiv thaiv kab mob, txawm tias cov tsos mob tseem pom nyob hauv pawg PRE raws li tau piav qhia yav dhau los [14]. Qhov kev soj ntsuam cuam tshuam thiab qhov hnyav ntawm cov kab mob hauv cov tsiaj no tsis raug ntsuas rau lub hom phiaj ntawm txoj kev tshawb no. Tus kab mob no tshwm sim txawv ntawm cov neeg laus thiab cov me nyuam mos uas muaj cov tsos mob hnyav dua thiab kev tuag hauv cov menyuam yug tshiab piv rau cov neeg laus [1,20]. Thaum lub sij hawm circulating neutralizing antibody qib ib leeg tsis muaj kev tiv thaiv [21], lawv yuav raug siv los ntsuam xyuas cov tshuaj tiv thaiv kab mob tom qab txhob txwm kis lossis txhaj tshuaj tiv thaiv hauv cov tsiaj. Thaum tsis muaj viremia yog tus yam ntxwv ntawm PEDV, viremia tau tshaj tawm hauv piglets txog 4 lub lis piam ntawm hnub nyoog [22,23]. Yog li, circulating neutralizing antibodies tuaj yeem cuam tshuam rau kev kho mob ntawm tus kab mob [21]. Txawm li cas los xij, kev tiv thaiv lactogenic, tshwj xeeb tshaj yog IgA hauv mammary secretions yuav yog qhov kev ntsuas ntau dua ntawm kev tiv thaiv piglets [21]. Cov kev ntsuam xyuas tau piav qhia hauv txoj kev tshawb no tuaj yeem siv los saib xyuas cov tshuaj tiv thaiv kab mob ntshav qab zib tom qab txhaj tshuaj tiv thaiv kab mob hauv cov pab pawg ntawm cov noob qoob loo lossis gilts ntxiv rau kev sau cov qog mammary los kuaj rau lactogenic / secretory IgA. Nws yuav muaj txiaj ntsig yog tias cov ntaub ntawv tsim tawm los ntawm cov kev ntsuam xyuas no tuaj yeem kwv yees qhov muaj nuj nqis thiab ruaj khov ntawm kev tiv thaiv lactogenic uas tsim los tom qab txhaj tshuaj tiv thaiv. Txawm li cas los xij, qhov ntawd tsis tau txheeb xyuas hauv qhov kev tshawb fawb tam sim no. Serum IgG cov tshuaj tiv thaiv kab mob tom qab txhaj tshuaj tiv thaiv tau pom tias yog kev kwv yees ntawm cov lus teb colostral IgG, tab sis tsis muaj kev sib raug zoo ntawm cov tshuaj tiv thaiv kab mob ntshav qab zib thiab cov qog mammary secretions [15]. Txawm hais tias tsis muaj kev sib raug zoo ntawm cov tshuaj tiv thaiv kab mob ntshav qab zib thiab cov kab mob mammary secretions, cov tsiaj txhaj tshuaj tau pom tias muaj cov tshuaj tiv thaiv kab mob siab dua hauv cov tshuaj tiv thaiv mammary piv rau cov tsiaj tswj [15]. Yog li ntawd, raws li kev txhawb nqa los ntawm, Bjustrom-Kraft, thiab al. [15], yog tias cov tshuaj tiv thaiv kab mob ntshav qab zib muaj siab, ces IgG thiab IgA hauv mammary secretions kuj yuav nce ntxiv. Yog li, kev sau cov ntshav kuaj ua ntej qhov kev txav ntawm cov poj niam mus rau lub tsev farrowing yuav txo qis kev ntxhov siab thiab mob tshwm sim thaum lub sij hawm periparturient sau cov ntshav thiab mammary secretions uas yuav cuam tshuam tsis zoo rau sow thiab litter [24]. Nws yog ib qho tseem ceeb rau cov neeg siv kom nkag siab tias tus nqi txiav tawm tuaj yeem cuam tshuam li cas rau kev kuaj mob rhiab heev thiab qhov tshwj xeeb ntawm qhov kev ntsuam xyuas. Kev nce tus nqi txiav tawm yuav txo qis cov txiaj ntsig tsis zoo thiab nce cov txiaj ntsig tsis zoo. Inversely, txo qis tus nqi txiav tawm yuav ua rau cov txiaj ntsig tsis zoo thiab txo cov txiaj ntsig tsis zoo. Qhov kev paub no tso cai rau tus neeg siv xaiv tus nqi txiav tawm los siv rau lawv qhov xwm txheej tshwj xeeb. Rau txoj kev tshawb no, raws li cov txiaj ntsig ntawm qhov kev tshawb fawb no thiab cov txiaj ntsig yav dhau los luam tawm los ntawm Sarmento li al. [2], peb xaiv 80 % FR rau HTNT thiab dilution ntawm 80 FFN. Ntawm cov kev txiav tawm kev kuaj mob tshwj xeeb yog siab dua rau HTNT (97%) piv rau FFN (76%) thiab muab cov txiaj ntsig zoo dua los ntawm HTNT piv rau FFN. Nws kuj yuav tsum to taub tias qhov kev kwv yees qhov tseem ceeb yog nyob ntawm qhov muaj kab mob ntau [25,26]. Nyob rau hauv txoj kev tshawb no, raws li cov qauv uas tau muab faib ua feem ntau yog 32.1% (36 qhov zoo thiab 76 tus qauv tsis zoo). Raws li kev nthuav dav zuj zus, PPV kuj tseem yuav nce ntxiv vim tias cov tsiaj kuaj (piv txwv) yuav zoo dua; Raws li prevalence txo, NPV yuav txo. Daim duab 3A thiab B qhia txog qhov no rau HTNT thiab FFN kev soj ntsuam ntawm ntau qhov kev txiav tawm uas tau soj ntsuam hauv qhov kev tshawb fawb no raws li cov kev piav qhia los ntawm Erb [25]. Yog li ntawd, thaum xav txog cov txiaj ntsig kev kuaj mob, ib tus yuav tsum xav txog qhov kev kuaj mob rhiab heev thiab qhov tshwj xeeb ntawm qhov kev ntsuam xyuas thiab nkag siab txog qhov muaj nyob hauv pab pawg yog siv cov txiaj ntsig kwv yees.
Daim duab 3. (A). Qhov txiaj ntsig zoo rau kev kwv yees rau HTNT (kab dub) thiab FFN (cov kab grey) kev soj ntsuam ntawm ntau qhov kev txiav tawm. Kev txiav tawm ntawm 90 tsis yog sawv cev rau HTNT. (B). Cov txiaj ntsig tsis zoo los yog HTNT (kab dub) thiab FFN (cov kab grey) kev soj ntsuam ntawm ntau qhov kev txiav tawm. Kev txiav tawm ntawm 20 tsis yog sawv cev rau FFN qhov kev xeem.
nws HTNT muaj kev kuaj mob zoo dua thiab qhov tshwj xeeb piv rau FFN. Thaum txheeb xyuas cov txiaj ntsig ntawm cov tshuaj tiv thaiv kab mob, qhov no yog qhov zoo tshaj plaws kom txo qis cov txiaj ntsig tsis zoo thiab cov txiaj ntsig tsis zoo. Piv txwv li, thaum siv kev txiav tawm ntawm 80 rau FFN, peb pom qhov rhiab heev ntawm 97% thiab qhov tshwj xeeb ntawm 76%. Qhov no txhais tau hais tias yuav muaj qee qhov tsis zoo tsis zoo tab sis yuav muaj ntau qhov tsis zoo. Yog tias cov txiaj ntsig tsis raug suav tias yog qhov zoo, qhov no yuav ua rau muaj kev txhais lus tsis tseeb tias ib pab tsiaj muaj cov tshuaj tiv thaiv rau PEDV thaum qhov tseeb lawv tsis ua. Txij li thaum HTNT muaj kev kuaj mob ntau dua thiab qhov tshwj xeeb thoob plaws ntau qhov kev txiav tawm, nws yog qhov kev ntsuam xyuas zoo dua rau kev kuaj pom ntawm PEDV cov tshuaj tiv thaiv. Cov txiaj ntsig los ntawm Sarmento et al. [2] tau pom ntau qhov kev kuaj mob ntau dua rau FFN thiab qhov sib piv rhiab heev thiab tshwj xeeb rau HTNT. Cov kev kuaj mob no tau pom muaj kev pom zoo raws li kappa statistic (κ). Ib qho tsis muaj zog ntawm κ yog tias ib qho kev xeem yuav tsum suav tias yog tus qauv kub. Thaum sib piv ob qhov kev ntsuam xyuas tsis zoo, qhov no tuaj yeem ua rau kev txhais lus ntawm κ. Rau txoj kev tshawb no, peb tau soj ntsuam qhov kev soj ntsuam kev ua tau zoo ntawm kev soj ntsuam nrog κ los txiav txim siab tias qhov kev ntsuam xyuas pom zoo tab sis tej zaum yuav muaj qhov sib txawv. Ib qho txiaj ntsig ntxiv ntawm HTNT dhau qhov kev ntsuam xyuas FFN yog tias HTNT cov txiaj ntsig tau suav los ntawm cov ntaub ntawv txuas ntxiv piv rau cov txiaj ntsig categorical (titrations) los ntawm FFN. Qhov xwm txheej tas mus li ntawm HTNT cov ntaub ntawv tso cai rau kev ntxub ntxaug ntau dua ntawm cov qauv ua piv txwv txhawm rau piav qhia txog qhov tsis zoo ntawm cov tshuaj tiv thaiv kab mob ntawm cov noob qoob loo. Raws li qhia los ntawm Sarmento et al. [2], HTNT muaj lub sijhawm nyeem luv luv ntawm 96 lub phaj zoo kom tsawg dua 4 min tso cai rau kev hloov pauv sai ntawm cov txiaj ntsig. Circulating systemic antibodies yuav pab tau piglet tiv thaiv kab mob PEDV [21]. Txawm li cas los xij, tsis muaj cov ntaub ntawv hais txog ntau npaum li cas ntawm cov tshuaj tiv thaiv kab mob hauv cov noob qoob loo yuav muab kev tiv thaiv PEDV rau piglets. Cov qib siab ntawm cov tshuaj tiv thaiv kab mob sib kis ua rau muaj ntau dua nyob rau hauv lub colostrum thiab muaj peev xwm tiv thaiv tau ntxiv. Kev ntsuam xyuas ntxiv tau lees paub los txiav txim siab txog kev sib koom ua ke ntawm, thaum lub sijhawm cev xeeb tub los sau cov qauv, thiab colostral antibody tiv thaiv kab mob PEDV. Lactogenic thiab mucosal tiv thaiv kab mob yog qhov tseem ceeb heev rau kev muaj sia nyob ntawm npua raug PEDV. Cov tshuaj tiv thaiv kab mob sib sau hauv cov mis ntawm lub plab-MG-sIGA axis thiab raug xa mus rau piglets kom muaj kev tiv thaiv. Langel et al. [27] pom tias gilts tshwm sim thaum lub sij hawm thib ob trimester muaj ntau dua ntawm kev ncig IgA, IgG, thiab cov tshuaj tiv thaiv tsis zoo piv rau thawj zaug thiab peb lub hlis twg gilts. Tsis tas li ntawd, Langel et al. [27] pom tias piglets ntawm gilts raug rau PEDV nyob rau hauv lub thib ob peb lub hlis twg muaj ib tug 100% ciaj sia taus tus nqi piv rau tsawg tshaj li 87.2% nyob rau hauv lwm lub trimesters qhia tias cov ntshav siab IgA, IgG, thiab neutralizing antibodies yog txuam nrog piglet ciaj sia nyob rau hauv lub ntsej muag ntawm PEB. Txawm hais tias cov tshuaj tiv thaiv kab mob ntshav qab zib tsis cuam tshuam txog kev tiv thaiv PEDV, cov txiaj ntsig los ntawm peb txoj kev tshawb fawb qhia tias cov tshuaj tiv thaiv kab mob ntshav qab zib muaj feem cuam tshuam nrog kev tiv thaiv kab mob PEDV. Raws li yav dhau los qhia los ntawm Brown li al. [14], PEDV raug tso rau hauv cov quav tom qab kev sib tw homologous thiab txhaj tshuaj tiv thaiv gilts. Txoj kev tshawb fawb tam sim no tau siv cov qauv los ntawm tib cov tsiaj li Brown li al. [14]. qhia ntxiv tias cov tshuaj tiv thaiv kab mob ntshav qab zib nce ntxiv tom qab muaj kev sib tw kis lossis txhaj tshuaj tiv thaiv tsis cuam tshuam rau tus qauv ntawm tus kab mob vim tias cov tshuaj tiv thaiv kab mob nce ntxiv tom qab kev sib tw thiab kev txhaj tshuaj. Cov kev tshawb fawb ntxiv tau lees paub los lees paub yog tias tus nqi ntawm tus kab mob tso rau hauv cov quav yog cuam tshuam nrog qhov muaj pes tsawg leeg ntawm cov tshuaj tiv thaiv kab mob uas muaj nyob hauv cov ntshav [21].
cistanche cov txiaj ntsig rau txiv neej-ua kom muaj zog tiv thaiv kab mob
5. Cov lus xaus
Lub null hypothesis ntawm txoj kev tshawb no yog tias tsis muaj qhov sib txawv ntawm qhov kev kuaj mob ntawm ob PEDV-neutralizing antibody assays, FFN thiab HTNT. Cov txiaj ntsig tau pom tias cov kev ntsuam xyuas no tau pom zoo raws li kappa statistic thaum muaj kev hloov pauv hauv lawv qhov kev kuaj mob thiab cov txiaj ntsig kwv yees raws li kev txiav tawm rau qhov kev xeem. Cov kev ntsuam xyuas no tuaj yeem siv los ntsuas qhov kev txhaj tshuaj tiv thaiv ntawm ib pab tsiaj. Kev tsis lees paub qhov kev xav tsis tseeb qhia tau hais tias HTNT yog qhov kev sim zoo tshaj plaws rau kev kuaj pom ntawm PEDV-neutralizing antibodies hauv cov ntshav. Tom qab txhaj tshuaj tiv thaiv hauv ob pawg PRE thiab POST, txoj kev tshawb no pom feem ntau ntawm cov txiaj ntsig zoo. Hauv pab pawg PRE, txo qis qhov sib txawv ntawm cov txiaj ntsig tau ua raws li qhov kev sib tw. Cov txiaj ntsig no qhia tau tias cov kws kho mob swine tuaj yeem ntsuas cov tshuaj tiv thaiv kab mob ntshav qab zib kom txiav txim siab seb cov tshuaj tiv thaiv PEDV puas raug tswj hwm zoo hauv pab tsiaj.
Cov ntaub ntawv
1. Saib, L.; Wang, Q.; Vlasova, UA; Jung, K.; Xiao, S. Coronaviruses. Hauv Kab Mob Swine, 11th ed.; Zimmerman, JJ, Ed.; Wiley Blackwell: Hoboken, NJ, USA, 2019.
2. Sarmento, LV; Poonsuk, K.; Tian, L.; Mora-Diaz, JC; Main, RG; Baum, DH; Zimmerman, JJ; Gimenez-Lirola, LG Kev Tshawb Fawb ntawm porcine kis kab mob raws plab kab mob-neutralizing antibody siv high-throughput imaging cytometry. J. Vet. Diagnos. Tshawb xyuas. 2020, 32, 324–328. [CrossRef]
3. Stevenson, GW; Hoang, H.; Schwartz, KJ; Burrough, ER; Sun, D.; Madson, D.; Cooper, VL; Pillatzki, UA; Gaug, P.; Schmitt, IB; ua al. Kev tshwm sim ntawm Porcine kab mob raws plab kab mob hauv Tebchaws Meskas: Cov tsos mob, qhov txhab, thiab kab mob kis kab mob. J. Vet. Diagnos. Tshawb xyuas. 2013, 25, 649–654. [CrossRef]
4. Kim, Y.; Yaj, M.; Goyal, SM; Cheeran, MC; Torremorell, M. Kev ntsuam xyuas ntawm biosecurity ntsuas los tiv thaiv kev kis tus kab mob porcine kab mob raws plab. BMC Vet. Res. 2017, 13, 89. [CrossRef]
5. Nkauj, DS; Aw, JS; Kang, BK; Yang, JS; hli, HJ; Yog, HS; Yaj, YS; Park, BK Qhov ncauj ua tau zoo ntawm Vero cell attenuated porcine epidemic diarrhea virus DR13 strain. Res. Vet. Sci. 2007, 82, 134–140. [CrossRef] [PubMed]
6. de Arriba, ML; Carvajal, UA; Pov, J.; Rubio, P. Mucosal thiab systemic isotype-specific antibody teb thiab kev tiv thaiv nyob rau hauv cov pa npua uas raug tus kab mob virulent los yog attenuated porcine kab mob raws plab kab mob. Vet. Immunol. Immunopathol. 2002, 85, 85–97. [CrossRef] [PubMed]
7. Furukawa, S.; Kuroda, Y.; Sugiyama, A. Kev sib piv ntawm cov qauv histological ntawm cov placenta hauv cov tsiaj sim. J. Toxicol. Pathol. 2014, 27, 11–18. [CrossRef] [PubMed]
8. Bourne, FJ; Curtis, J. Kev hloov ntawm immunoglobins IgG, IgA, thiab IgM los ntawm cov ntshav mus rau colostrum thiab mis nyuj hauv cov noob. Immunology 1973, 24, 157–162. [PubMed]
9. Parren, PW; Burton, DR Cov tshuaj tiv thaiv kab mob ntawm cov tshuaj tiv thaiv kab mob hauv vitro thiab hauv vivo. Adv. Immunol. 2001, 77, 195–262. [CrossRef]
10. Bjustrom-Kraft, J.; Woodard, K.; Gimenez-Lirola, L.; Rotolo, M.; Wang, C.; Sun, Y.; Lasley, P.; Zhang, J. Baum, D.; Gaug, P.; ua al. Porcine epidemic diarrhea virus (PEDV) kuaj pom thiab tshuaj tiv thaiv kab mob hauv cov npua ua lag luam. BMC Vet. Res. 2016, 12, 99. [CrossRef]
11. Aw, JS; Nkauj, DS; Yang, JS; Nkauj, JY; hli, HJ; Kim, TY; Park, BK Kev sib piv ntawm enzyme-linked immunosorbent assay nrog kev kuaj ntshav qabzib rau serodiagnosis ntawm porcine epidemic diarrhea kab mob. J. Vet. Sci. 2005, 6, 349–352. [CrossRef]
12. Okda, F.; Li, X.; Singrey, UA; Clement, T. Nelson, J.; Christopher-Hennings, J.; Nelson, EA; Lawson, S. Kev loj hlob ntawm kev tsis ncaj ncees ELISA, thaiv ELISA, fluorescent microsphere immunoassay thiab fluorescent tsom mus rau qhov kev ntsuam xyuas tsis zoo rau kev soj ntsuam serologic ntawm kev raug rau North American hom kab mob Porcine Epidemic Diarrhea Virus. BMC Vet. Res. 2015, 11, 180. [CrossRef] [PubMed]
13. Chen, Q.; Li, G.; Stasko, J.; Thomas, JT; Stensland, WR; Pillatzki, AE; Gauge, PC; Schwartz, KJ; Madson, D.; Yoon, KJ; ua al. Kev cais tawm thiab ua tus cwj pwm ntawm cov kab mob porcine kab mob raws plab kab mob cuam tshuam nrog 2013 kab mob tshwm sim ntawm swine hauv Tebchaws Meskas. J. Clin. Microbiol. 2014, 52, 234–243. [CrossRef]
14. Brown, J.; Rademacher, C.; Baker, S. Kev ua tau zoo ntawm kev lag luam porcine kis kab mob raws plab kab mob tshuaj tiv thaiv ntawm kev txo lub sijhawm ntawm tus kab mob hauv cov gilts. JSHAP 2019, 27, 256–264.
15. Bjustrom-Kraft, J.; Woodard, K.; Giménez-Lirola, L. Serum thiab mammary secretion antibody teb nyob rau hauv porcine epidemic diarrhea-immune gilts tom qab porcine kis kab mob raws plab txhaj tshuaj tiv thaiv. JSHAP 2018, 26, 34–40.
16. Iowa State University. Cov txheej txheem rau Porcine Epidemic Diarrhea Virus (PEDV) Fluorescent Focus Neutralization (FFN) Test. Serology 2017, 1–7.
17. Iowa State University. Cov txheej txheem rau Porcine Epidemic Diarrhea Virus (PEDV) High-Throughput Virus Neutralization Test (HTNT). Serology 2017, 1–16.
18. Hierholzer, JC; Killington, RA; Kangro, HO; Mahy, BWJ Virology Methods Manual; Kev Kawm Xov Xwm: London, UK, 1996; p. 21.
19. Thrusfield, M. Veterinary Epidemiology, 2nd ed.; Blackwell Science LTD: Hoboken, NJ, USA, 1995; p. 479 ib.
20. Lee, C. Porcine epidemic diarrhea virus: Ib tus kab mob uas tshwm sim thiab rov tshwm sim epizootic swine virus. Virol. J. 2015, 12, 193. [CrossRef]
21. Poonsuk, K.; Gimenez-Lirola, LG; Zhang, J. Arruda, P.; Chen, Q.; Correa da Silva Carrion, L. Magtoto, R.; Pineyro, P.; Sarmento, L.; ib. Wang, C.; ua al. Puas Circulating Antibody Ua lub luag haujlwm hauv kev tiv thaiv Piglets tiv thaiv tus kab mob Porcine Epidemic Diarrhea? PLoS ONE 2016, 11, e0153041. [CrossRef]
22. Niederwerder, MC; Nietfeld, JC; Bai, J.; Peddireddi, L.; Breazale, IB; Anderson, J.; Kerrigan, MA; Ib, B.; Oberst, RD; Crawford, K.; ua al. Cov ntaub so ntswg hauv zos, tso tawm, kev thauj tus kab mob, cov tshuaj tiv thaiv kab mob, thiab kev xa cov aerosol ntawm Porcine kab mob raws plab kab mob tom qab inoculation ntawm 4-lub lim tiam-laus feeder npua. J. Vet. Diagnos. Kev nqis peev. 2016, 28, 671–678. [CrossRef]
23. Madson, DM; Arruda, PH; Magstadt, DR; Burrough, ER; Hoang, H.; Sun, D.; Bower, LP; Bhandari, M.; Gauge, PC; Stevenson, GW; ua al. Tus cwj pwm ntawm Porcine Epidemic Diarrhea Virus Isolates US/Iowa/18984/2013 Infection in 1-Hnub-laus Cesarean-Derived Colostrum-Deprived Piglets. Vet. Pathol. 2016, 53, 44–52. [CrossRef]
24. Maiau, E.; Manteca, X. Mob thiab tsis xis nyob los ntawm kev yug me nyuam hauv nyuj thiab tseb. Appl. Anim. Behav. Sci. 2011, 135, 241–251. [CrossRef]
25. Erb, HN Qhov tshwm sim ua ntej (qhov kev kwv yees zoo tshaj plaws ua ntej) cuam tshuam cov txiaj ntsig ntawm kev kuaj mob. Vet. Clin. Pathol. 2011, 40, 154–158. [CrossRef] [PubMed]
26. Tenny, S.; Hoffman, MR Prevalence. Hauv StatPearls; StatPearls Publishing: Treasure Island, FL, USA, 2022. 27. Langel, SN; Paim, FC; Alhamo, MA; Buckley, ib.; Van Geelen, A.; Lager, KM; Vlasova, UA; Saif, LJ Theem Gestation at Porcine Epidemic Diarrhea Virus Infection of Pregnant Swine Impacts Maternal Immunity and Lactogenic Immune Protection of Neonatal Suckling Piglets. Pem hauv ntej. Immunol. 2019, 10, 727. [CrossRef] [PubMed]