检测猪流行性腹泻病毒、猪传染性胃肠炎病毒和猪轮状病毒的cDNA芯片的构建

doi:10.11843/j.issn.0366-6964.2015.12.015

Abstract

Abstract:

Porcine Epidemic Diarrhea Virus(PEDV)，Transmissible Gastroenteritis Virus(TGEV) and Porcine Rotavirus(PoRV) are three common pathogenies causing piglets diarrhea.A novel cDNA microarray was necessary to be developed to simultaneous detection of three diarrheal viruses successfully.To make this possible，primers and probes were designed to amplification of target genes according to the conserved sequences of PEDV-S，PEDV-M，TGEV-S，TGEV-N，PoRV-VP7 and PoRV-NSP4.Mutiplex PCR was established for labeling target genes after designing the microarray matrix and finally evaluated the specificity，sensitivity and repeatability of microarray.The positive judgement standard of microarray was determined：SNR(Signal to Noise Ratio) ≥2.0 or median signal ≥1 500.The result showed that PRRSV，CSFV，JEV and PRV could not be detected by six probes and as few as 20 pg•μL^-1 of target plasminds were detected successfully.One microarray could be tested at least 7 times.This assay provided a novel and high throughput detecting technology to identification and detection of three diarrheal viruses.

CLC Number:

HUA Xiang，HU Zhong-kai，HUANG Xiao-bo，WEN Xin-tian，CAO San-jie，WEN Yi-ping，WU Rui，DENG Jing，ZHAO Song，YIN Ren-jie，CHANG Xiao-xia，OUYANG Da，ZHANG Xian. Construction of cDNA Microarray for Detecting Porcine Epidemic Diarrhea Virus，Porcine Transmissible Gastroenteritis Virus and Porcine Rotavirus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(12): 2235-2242.

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Construction of cDNA Microarray for Detecting Porcine Epidemic Diarrhea Virus，Porcine Transmissible Gastroenteritis Virus and Porcine Rotavirus

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