Development and testing of procedures for detecting HPV 6 and HPV 16 DNA in water samples

Abstract

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.