Development of loop-mediated isothermal amplification assay for specific and rapid detection of differential goat pox virus and sheep pox virus.
Development of loop-mediated isothermal amplification assay for specific and rapid detection of differential goat pox virus and sheep pox virus.
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开发用于特异性快速检测差异山羊痘病毒和绵羊痘病毒的环介导等温扩增试验
DOI：
10.1186/1471-2180-14-10
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发表时间：
2014-01-17
期刊：
影响因子：
4.2
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Zhao Z;Fan B;Wu G;Yan X;Li Y;Zhou X;Yue H;Dai X;Zhu H;Tian B;Li J;Zhang Q
羊痘病毒是世界上山羊和绵羊产区重要的经济病原体，主要包括山羊痘病毒(GTPV)、绵羊痘病毒(SPPV)和块状皮肤病病毒(LSDV)。临床上，羊痘和羊痘具有相同的症状，在血清学上无法区分。这就需要一种快速、廉价、易于操作和维护的基因分型工具，以便于准确的疾病诊断和监测，以便更好地管理羊痘暴发。结果根据ITR序列设计了三套LAMP引物，建立了一种用于GTPV和SPPV特异性鉴别检测的LAMP方法。反应在62℃下进行45或60分钟，并通过对几个GTPV和SPPV分离株的成功差异检测证实了特异性。与口蹄疫病毒(FMDV)、边缘绿僵菌、支原体亚种无交叉反应。对135份保存的疫情材料进行RFLP-PCR分析，羊痘48份，羊痘87份，LAMP检测结果均为阳性。当利用GTPV和SPPV基因组DNA时，通用LAMP和GTPV LAMP引物的检出率为100%，而SPPV LAMP的检出率为98.8%，与实验室检测结果一致。结论三套LAMP引物组合使用，提供了一种分析可靠的方法，能够完全区分GTPV和SPPV。LAMP方法为区分GTPV和SPPV感染提供了一种特异、敏感、快速的诊断工具，有可能作为流行地区羊痘病毒的一种标准化检测方法。
BackgroundCapripox viruses are economically important pathogens in goat and sheep producing areas of the world, with specific focus on goat pox virus (GTPV), sheep pox virus (SPPV) and the Lumpy Skin Disease virus (LSDV). Clinically, sheep pox and goat pox have the same symptoms and cannot be distinguished serologically. This presents a real need for a rapid, inexpensive, and easy to operate and maintain genotyping tool to facilitate accurate disease diagnosis and surveillance for better management of Capripox outbreaks.ResultsA LAMP method was developed for the specific differential detection of GTPV and SPPV using three sets of LAMP primers designed on the basis of ITR sequences. Reactions were performed at 62°C for either 45 or 60 min, and specificity confirmed by successful differential detection of several GTPV and SPPV isolates. No cross reactivity withOrfvirus, foot-and-mouth disease virus (FMDV),A. marginale Lushiisolate, Mycoplasma mycoides subsp. capri, Chlamydophila psittaci, Theileria ovis, T. luwenshuni, T. uilenbergiorBabesia spwas noted.RFLP-PCR analysis of 135 preserved epidemic materials revealed 48 samples infected with goat pox and 87 infected with sheep pox, with LAMP test results showing a positive detection for all samples. When utilizing GTPV and SPPV genomic DNA, the universal LAMP primers (GSPV) and GTPV LAMP primers displayed a 100% detection rate; while the SPPV LAMP detection rate was 98.8%, consistent with the laboratory tested results.ConclusionsIn summary, the three sets of LAMP primers when combined provide an analytically robust method able to fully distinguish between GTPV and SPPV. The presented LAMP method provides a specific, sensitive and rapid diagnostic tool for the distinction of GTPV and SPPV infections, with the potential to be standardized as a detection method for Capripox viruses in endemic areas.
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影响因子：
1.6
作者：
Hosamani, M;Mondal, B;Rasool, TJ
通讯作者：
Rasool, TJ
影响因子：
3.8
作者：
GERSHON, PD;KITCHING, RP;BLACK, DN
通讯作者：
BLACK, DN
影响因子：
3.3
作者：
Lamien, Charles Euloge;Le Goff, Christian;Diallo, Adama
通讯作者：
Diallo, Adama
影响因子：
4.2
作者：
KITCHING, RP;BHAT, PP;BLACK, DN
通讯作者：
BLACK, DN
中国甘肃省爆发与山羊痘病毒相关的绵羊痘疫情
影响因子：
3.3
作者：
Yan, Xin-Min;Chu, Yue-Feng;Zhang, Qiang
通讯作者：
Zhang, Qiang