南方农业学报2011，Vol.42Issue(12)：1545-1549,5.
TaqMan-LNA探针荧光定量PCR快速检测对虾传染性皮下及造血组织坏死病毒
Rapid detection of infectious hypodermal and hematopoietic necrosis virus in shrimps using TaqMan-LNA probe real-time PCR
摘要
Abstract
[Objective]The present experiment was conducted to develop a TaqMan-LNA probe real-time PCR assay for detection of infectious hypodermal and hematopoietic necroses virus (IHHNV) to provide convenient detection technology for clinical diagnosis and epidemic monitoring of IHHNV. [Method] The specific primers and TaqMan-LNA probes were designed from conserved IHHNV sequences obtained from GenBank. The real-time PCR assay was optimized to reduce the detection time and to improve the sensitivity and specificity. Three hundred clinical shrimp samples collected from Cuangxi were detected by real-time PCR to evaluate practicality of assay. [Result]The real-time PCR assay accurately detected and quantified IHHNV. It showed better linear relationship from 1.6×101 to 1.6×l08 copies/μL and had a minimum detection limit of about 10 virion. The method was specific for IHHNV and had no cross-reaction with three other shrimp pathogen such as white spot syndrome virus (WSSV), Vibrio parahaemolyticus, taura syndrome virus (TSV). This method also showed excellent reproducibility with the coefficient of variations ranging from 0.53-1.75% within group and 0.81-1.14% among different groups. Out of 300 samples, 193 (64.3%) showed positive results for IHHNV which revealed that IHHNV infection in Litopenaeus vannanei is a major threat for shrimp cultivation in Guangxi. [Conclusion]The TaqMan-LNA probe real-time PCR assay is a quick, sensitive, specific, quantitative and reproducible tool for detecting IHHNV.关键词
凡纳滨对虾/传染性皮下及造血组织坏死病毒(IHHNV)/TaqMan-LNA探针/荧光定量PCRKey words
Litopenaeus vannanei/ IHHNV/ TaqMan-LNA probe/ real-time PCR分类
农业科技引用本文复制引用
韦信贤,童桂香,谢宗升,吴祥庆,黄国秋,黄玉柳,廖永志,黎小正..TaqMan-LNA探针荧光定量PCR快速检测对虾传染性皮下及造血组织坏死病毒[J].南方农业学报,2011,42(12):1545-1549,5.基金项目
广西直属公益性科研院所基本科研业务费专项项目(2060302GXIF-2009-05) （2060302GXIF-2009-05）
