Comparison of Different PCR Tests to Detect Bovine Lentivirus in Cell Culture and Experimentally and Naturally Infected Cattle
Comparison of Different PCR Tests to Detect Bovine Lentivirus in Cell Culture and Experimentally and Naturally Infected Cattle
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细胞培养物以及实验和自然感染牛中牛慢病毒不同 PCR 检测方法的比较
DOI：
10.1177/104063879700900415
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发表时间：
1997
影响因子：
1.5
通讯作者：
C. Whetstone
中科院分区：
文献类型：
--
作者：
D. Suarez;C. Whetstone
靶基因反应体积DNA引物dNTPs Taq聚合酶MgCl2循环热启动Misc。env,波尔50毫升0.5毫克200毫米1.25单位3毫米pmol嵌套- 62周期没有波尔和插科打诨50毫升2.0毫克25 pmol 200毫米2.5单位1.5毫米40周期是的0.01%明胶0.1% Triton x - 100波尔100毫升5000细胞10 pmol 200 mM 5单位1.5毫米35周期是的牛慢病毒,也被称为牛immunodeficiency-like病毒(出),会持续的病毒感染牛,但是目前,它是未知的,如果导致牛临床疾病或临床免疫缺陷。只有一项关于BIV的流行病学研究报告，该研究将奶牛产奶量下降与BIV联系起来。4为了测试BIV是否会导致牛患病，确定牛是否感染BIV的可靠诊断测试至关重要。一个好的诊断测试应该是特异性的、敏感的、廉价的，并能快速提供结果。然而，目前描述的BIV测试没有一个符合所有这些类别。血清学检测是乙肝病毒最常用的诊断检测方法。然而，有报道称，在持续感染BIV的实验感染牛中，对BIV蛋白的抗体丢失p26 gag抗原（western blot试验和一些ELISA试验中用于检测BIV的最重要抗原）抗体的缺失可能导致动物被认为未感染该病毒，即使通过PCR和病毒分离均可证实该病毒8,10,15从自然感染的牛身上分离BIV病毒仍然很困难，仅报道了3例分离病毒2、11、13在其他病毒系统中，用PCR进行病毒诊断已被证明是一种敏感的诊断测试，尽管已经发表了5种用于检测BIV的PCR测试，但它们的敏感性和特异性尚未进行比较本文的目的是对目前描述的PCR检测方法进行比较，并对一些PCR条件进行标准化。三份不同的报告共描述了5种不同的PCR检测（表1）在所有5项测试中，总共使用了39种不同的模板，每次反应0.5 mg DNA（表2）
Genes targeted Reaction volume DNA Primers dNTPs Taq polymerase MgCl2 Cycles Hot start Misc. env and pol 50 ml 0.5 mg 20 pmol 200 mM 1.25 units 3 mM nested—62 cycles no pol and gag 50 ml 2.0 mg 25 pmol 200 mM 2.5 units 1.5 mM 40 cycles yes 0.01% gelatin 0.1% Triton X-100 pol 100 ml 5,000 cells 10 pmol 200 mM 5 units 1.5 mM 35 cycles yes The bovine lentivirus, also known as the bovine immunodeficiency-like virus (BIV), causes a persistent viral infection in cattle, but at present, it is unknown if it causes clinical disease or clinical immunodeficiency in cattle. Only a single epidemiological study has been reported with BIV, and this study associated a decreased milk yield in cows with BIV.4 In order to test whether BIV causes disease in cattle, a reliable diagnostic test to determine if cattle are infected with BIV is critical. A good diagnostic test should be specific, sensitive, and inexpensive and provide results quickly. However, none of the currently described BIV tests fit all of these categories. Serological tests have been the most commonly used diagnostic tests for BIV. However, loss of antibody to BIV proteins in experimentally infected cattle that are persistently infected with BIV has been reported.3,7,9,10,14 The loss of antibody to the p26 gag antigen, the most important antigen for detection of BIV in western blot tests and in some ELISA tests, can result in an animal considered to be uninfected with this virus even when virus can be demonstrated by both PCR and virus isolation.8,10,15 Virus isolation of BIV from naturally infected cattle remains difficult, with only 3 reported virus isolations.2,11,13 Viral diagnosis by PCR has been shown to be a sensitive diagnostic test in other viral systems, and although 5 PCR tests have been published for the detection of BIV, their sensitivity and specificity have not been compared.5,6,10 The purpose of this paper is to provide a comparison among the currently described PCR tests with standardization of some of the PCR conditions. Three different reports describe a total of 5 different PCR tests (Table 1).5,6,10 For all 5 tests, a total of 39 different templates were used with 0.5 mg of DNA per reaction (Table
通过在实验感染的小牛中进行巢式聚合酶链反应测试，改进了牛慢病毒的早期和长期检测。
DOI：
--
发表时间：
1995
期刊：
American journal of veterinary research.
影响因子：
--
作者：
Suarez,DL;VanderMaaten,MJ;Whetstone,CA
通讯作者：
Whetstone,CA
影响因子：
3.7
作者：
D. Suarez;C. Whetstone
通讯作者：
D. Suarez;C. Whetstone