开发和测试水样中HPV 6和HPV 16 DNA检测程序

A. Stolbikov, R. Salyaev, N. Rekoslavskaya
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引用次数: 1

摘要

本研究旨在开发和测试检测水样中危险的人类乳头瘤病毒(HPV) 6型和16型DNA的程序。在NCBI (National Center for Biotechnology Information)数据库和BioEdit程序的帮助下,采用生物信息学方法研究了HPV 6l1和HPV 16l1核酸序列的保守片段。共检测了135个HPV6 L1核酸序列和945个HPV16 L1核酸序列。利用专用程序(PerlPrimer v.1.1.21、FastPCR 6.6和Primer3Plus)对鉴定出的核酸序列保守片段开发5对特异性引物。此外,还测试了从Listvyanka定居点(贝加尔湖)附近的各种水体中收集样本的几种程序。对样品进行不溶性颗粒和细菌污染的全面纯化,利用与HPV6 L1和HPV16 L1核酸序列互补的引物进行PCR分析,检测HPV DNA的存在。所进行的研究显示,水样中含有HPV 6和HPV 16 DNA。由于使用了开发和测试的程序,从贝加尔湖自然领土的各种水源收集和检查样本,然后进行聚合酶链反应分析,因此有可能检测到危险病毒的存在。拟议的检测水样是否存在HPV的程序可用于对贝加尔湖和其他地区的水体和废水进行有效监测。
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Development and testing of procedures for detecting HPV 6 and HPV 16 DNA in water samples
The present study aims to develop and test procedures for detecting the DNA of dangerous human papillomaviruses (HPV) types 6 and 16 in water samples. The conserved segments of HPV 6 L1 and HPV 16 L1 nucleic acid sequences were studied using bioinformatic methods with the help of the NCBI (National Center for Biotechnology Information) database and the BioEdit program. A total of 135 nucleic acid sequences of HPV6 L1 and 945 nucleic acid sequences of HPV16 L1 were examined. Five pairs of specific primers were developed for the identified conserved segments of nucleic acid sequences using specialized programs (PerlPrimer v.1.1.21, FastPCR 6.6, and Primer3Plus). In addition, several procedures for collecting samples from various water bodies located near Listvyanka settlement (Lake Baikal) were tested. The samples were subjected to comprehensive purification from insoluble particles and bacterial contamination to be tested for the presence of HPV DNA via PCR analysis using primers complementary to the nucleic acid sequences of HPV6 L1 and HPV16 L1. The conducted studies revealed HPV 6 and HPV 16 DNA in the water samples. Due to the use of the developed and tested procedures for collecting and examining samples from various water sources in the Baikal Natural Territory followed by a PCR analysis, it was possible to detect the presence of dangerous viruses. The proposed procedure of testing water samples for the presence of HPV can be useful in developing effective monitoring of water bodies and wastewater both in Baikal and other regions.
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