Cov kab laum Nyom yog ib qho tseem ceeb nyiaj txiag hauv Tuam Tshoj cov dej afaculture epaculture. Txawm li cas los xij, nyob rau hauv xyoo tsis ntev los no, tusHepatopaleatic Microsampium(EHP), Ib tug parasite kis tus kab mob hepatopenas, muaj kev cuam tshuam loj heev rau kev lag luam. Vim tias tam sim no tsis muaj tshuaj tiv thaiv cov tshuaj muaj txiaj ntsig los kho tus kab mob no, kev paub sai thiab yog qhov tseem ceeb heev los tiv thaiv nws kis.
A.Magnified 9. {{0 × 1 {{4 {4}} 3 zaug; B. MAGTIFIED 5.0 × 104 zaug.
Qiao yi, Shen hui, wan xihe, ntxuam xianping, jiang ge, li hui, wang libao, shi wenjun, cheng Jie. Kev rho tawm thiab kev soj ntsuam morphological ntawm Hepatopancreat Microsporidium hauv Litropteidium Vannamei. Phau ntawv sau xov xwm ntawm Fishery Sciences ntawm Tuam Tshoj, 2018, 25 (5): 1051-1058. DOI: 10.3724 / sp.j.1118.2018.17430.
Kev kuaj pom ntawm EHP feem ntau txoj kev vam meej ntawm molecular txoj kev biology, nrog PV technology yog cov nquag tshaj plaws. Txawm li cas los xij, thaum PCR muaj cov siab rhiab siab, nws yuav tsum muaj cov cuab yeej PC cov khoom siv tshwj xeeb, ua rau nws tsis ua haujlwm rau kev siv. Txhawm rau daws cov teebmeem no, cov pab pawg tshawb fawb tau tsim ob txoj hauv kev nrawm nrawm raws li MMA thev naus laus zis. Cov hau kev no muaj cov lus teb siab, cov lus teb sai, ua haujlwm yooj yim, yog li sau cov khoom siv ntawm EHP hauv cov kev tshawb pom cov kev kuaj pom tam sim no.
Amplification reagents siv nyob rau hauv kev kawm:
- DNA yog nthuav tawm cov khoom siv sai sai sai (theem pib)
- DNA yog nthuav tawm cov khoom siv sai sai (colloidal kub sawb hom)
Mira Technology yog ntau-enzyme yog Ootothermal nrawm nucleic acid didification txoj kev. Nws cov ntsiab lus tseem ceeb koom nrog txoj kev ua haujlwm ntawm ntau cov protein abient ua kom sov ua kom sov, ua rau lub ntsej muag zoo nkauj, rau ntawm nucleic acid kuaj tau.
Kev Tshawb Fawb
Qauv extractor
Pab neeg tshawb fawb tau sau ob tus kab mob thiab kev noj qab haus huv cov qauv los ntawm peb lub xeev shrimp hauv lub xeev ntawm lub nroog Hepatopleas.
Qhov tsim thawj tsim qauv
Peb teev ntawm MMA Primers tau tsim cov hom phiaj S8 serine protein gene ntawm EHP (Genbank Access Access Dance: ntawm 182063). Cov noob no muaj cov chaw ua haujlwm tshwj xeeb, kom muaj feem tshwj xeeb ntawm kev kuaj pom.
Mira-Hnub nyoog thiab Mira-LFD nrhiav kom tau
(A) Optimized RPA-hnub nyoog uas cov tshuaj tiv thaiv kub. M: DL 500 tus cim; N: kev tswj hwm tsis zoo; Cov kab mob 1-5: 25 degree, 30 degree, 37 degree, 39 degree, thiab 42 degree.
(B) cov hnub nyoog rpa-hnub nyoog. M: DL 500 tus cim; N: kev tswj hwm tsis zoo; 1: 10 min; 2: 20 min; 3: 30 min; 4: 40 min.
(C) Optimized RPA-LFD cov tshuaj tiv thaiv kub. N: kev tswj hwm tsis zoo; 1: 25 degree; 2: 30 degree; 3: 37 degree; 4: 39 degree; 5: 42 Qib.
(D) Optimized RPA-LFD cov tshuaj tiv thaiv. N: kev tswj hwm tsis zoo; 1: 5 min; 2: 10 min; 3: 20 min.
Ob txoj kev pom ntawm Mira thiab muaj hnub nyoog Mira, agarose gel electrophoresis) thiab Mira-LFD (colloidal Kub Forf Flatoassay), tau siv los kuaj EHP. Pab neeg tshawb fawb txog cov tshuaj tiv thaiv huab cua thiab sijhawm sim. Mira-lub hnub nyoog nthuav tawm DNA hauv 30 feeb ntawm 37 degree, thaum Mira-LFD ua tiav kev ua tiav hauv 107 degree, nrog cov txiaj ntsig tau nyeem tau hauv 5 feeb.
Sim cov txiaj ntsig
Tshwj xeeb thiab rhiab
(A) tshwj xeeb ntawm RPA-etection kev paub. M: DL 1000 cim; P: nosema bombycis; 1: EHP; 2: encephalationozoon; 3: microstroridia; 4: Dawb Spordrrome tus kab mob; 6: Txawv txawv; N: kev tswj hwm tsis zoo.
(B) tshwj xeeb ntawm RPA-LFD tshawb nrhiav. P: nosema bombycis; N: kev tswj hwm tsis zoo; 1: EHP; 2: encephalationozoon; 3: microstroridia; 4: Dawb Spordrrome tus kab mob; 6: Txawv txawv.
(C) rhiab ntawm RPA-lub hnub nyoog nrog kev ua kom tiav ntawm cov txiaj ntsig zoo ntawm cov plasmids. M: DL 500 tus cim; N: kev tswj hwm tsis zoo; 1: 4.7 × 10 ^ 5 daim ntawv / μLE; 2: 4.7 × 10 ^ 4 daim ntawv luam / μl; 3: 4.7 × 10 ^ 3 daim ntawv / μLE; 4: 4.7 × 10 ^ 2 daim ntawv luam / μl; 5: 4.7 Luam / μl; 6: 4.7 Luam / μl.
(D) Cov rhiab ntawm RPA-LFD nrog cov txiaj ntsig gradient ntawm cov plasmids rov qab. N: kev tswj hwm tsis zoo; 1: 4.7 × 10 ^ 5 daim ntawv / μLE; 2: 4.7 × 10 ^ 4 daim ntawv luam / μl; 3: 4.7 × 10 ^ 3 daim ntawv / μLE; 4: 4.7 × 10 ^ 2 daim ntawv luam / μl; 5: 4.7 Luam / μl; Thiab 6: 4.7 Luam / μl.
Qhov kev sim ua tau pom tias Mira Technology muaj qhov tshwj xeeb, kuaj pom tseeb EHP yam tsis muaj lwm yam pathogens. Nyob rau hauv cov nqe lus ntawm rhiab heev, ob txoj hauv kev muaj kev txwv tsis pub muaj 4.7 luam / μLE siab dua qhov rhiab heev ntawm pcr.
Cov cuab soj ntsuam kuaj xyuas
Cov txiaj ntsig los ntawm kev kuaj 30 chaw kuaj mob pom tau tias Mira-LFD muaj qhov zoo tshaj QPRCR's 80% tab sis siab dua li cov pristic 56.7%.
Thov qhuas
Mira-based cov qauv siv tau tso cai rau kev ua kom sai ntawm cov hom phiaj ntawm qhov kub tsis tu ncua. Tshwj xeeb, cov qauv mira-LFD tuaj yeem xa cov khoom siv tau 10 feeb yam tsis muaj kev xav tau rau kev siv cov liaj teb cws. Lub siab rhiab heev thiab qhov tshwj xeeb ntawm cov thev naus laus zis ua nws cov cuab yeej nrhiav thaum ntxov thiab kev tiv thaiv kev noj qab haus huv thiab kev txhim kho ntawm cov nyom cws ua liaj ua teb.
Ntawm cov hau kev no, colloidal Kub tom qab ndlub mouldoassay (Mira-LFD) yog qhov tshwj xeeb rau cov ntaub ntawv ntawm cov xaib kom yooj yim thiab cov txiaj ntsig yooj yim. Cov nyom ntoo cov nyom tuaj yeem tshawb pom thiab teb tus mob EHP sai sai, txhim kho kev ua liaj ua teb tau zoo, thiab txo cov nyiaj noj muaj txiaj ntsig.