Comparison of nested PCR and real-time PCR for the detection of Toxoplasma gondii in biological samples from naturally infected cats
Comparison of nested PCR and real-time PCR for the detection of Toxoplasma gondii in biological samples from naturally infected cats
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DOI：
10.1016/j.rvsc.2010.02.020
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发表时间：
2010-10-01
影响因子：
2.4
通讯作者：
Fuentes, Isabel
中科院分区：
文献类型：
--
作者：
Montoya, Ana;Miro, Guadalupe;Fuentes, Isabel
有不同的分子诊断方法，包括DNA提取方法来诊断弓形虫，有必要在生物样品中进行比较研究。本研究旨在比较实时荧光PCR（rtPCR）和巢式PCR（nPCR）检测T。自然感染的猫对弓形虫阳性猫的生物标本进行弓形虫检测。弓形虫DNA-通过裂解缓冲液和蛋白酶K（LB蛋白酶K）方法和酸性硫氰酸胍（GuSCN）方法提取-使用rtPCR和nPCR。t.用LB蛋白酶K法和GuSCN法提取弓形虫DNA，nPCR检出率分别为43.6%和40.8%。使用rtPCR，这些数字显著下降至33.8%和14.1%。尽管nPCR显示出更高的灵敏度，但两种PCR之间观察到的一致性良好;然而，这种一致性受到所使用的DNA提取方法的影响，LB蛋白酶K法显示出更好的结果。(C)2010爱思唯尔有限公司保留所有权利。
There are different protocols of molecular diagnosis methods available including DNA extraction methods to diagnose of Toxoplasma gondii, being necessary to perform comparative studies in biological samples. The aim of this study is to compare real-time PCR (rtPCR) and nested PCR (nPCR) to evaluate the detection of T. gondii in naturally infected cats. Biological samples of Toxoplasma-seropositive cats were assayed for detection of T. gondii DNA - extracted by both the lysis buffer and proteinase K (LB proteinase K) method and the acid guanidinium thiocyanate (GuSCN) method - using rtPCR and nPCR. T. gondii DNA was detected by nPCR in 43.6% and 40.8% of the samples from which it was extracted by the LB proteinase K and the GuSCN method, respectively. With rtPCR these figures fell significantly to 33.8% and 14.1%. Despite of nPCR showed higher sensitivity, the agreement observed between two PCRs was good; this agreement, however, was affected by the DNA extraction method used, LB proteinase K method showed better results. (C) 2010 Elsevier Ltd. All rights reserved.