Лабораторная диагностика гриппа типов A и Б методом ПЦР
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Ключевые слова:
вирус гриппа, ПЦР, специфические праймеры, амплификация, специфичность, чувствительность.Аннотация
В настоящей работе представлен разработанный нами метод для выявления РНК вируса гриппа типов А и Б c использованием мультипраймерной полимеразной цепной реакции. Подобраны специфичные праймеры – InfАM63 и InfАM258, характерные для вируса гриппа типа А и специфичные праймеры InfBM26 и InfBM293, характерные для вируса гриппа типа Б. Специфичность полимеразной цепной реакции была протестирована на нескольких штаммах вируса гриппа типов А и Б, полученных из коллекции микроорганизмов НИИПББ КН МОН РК. Высокая специфичность тест-системы обусловлена тем, что в исследуемом материале выявляется уникальный, характерный только для вируса гриппа типов А и Б фрагмент РНК. Чувствительность разработанной полимеразной цепной реакции определена путем проведения реакции с различными разведениями вирусной РНК. Порог чувствительности вируса гриппа типов A и Б составляет 1х102 копий РНК (0,1 пг) в пробе. Диагностика вируса гриппа типов А и Б c использованием полимеразной цепной реакции позволяет точно и очень быстро (~5-6 часов) выявлять РНК вируса гриппа типов А и Б из клинического материала. При этом возможно одновременное исследование РНК вируса гриппа на типы А и Б.
Библиографические ссылки
1 Quilivan M., Cullinane A., Nelly M., et al. Comparison of Sensitivities of Virus Isolation, Antigen Detection and Nucleic Acid Amplification for Detection of Equine Influenza Virus // J. Clin. Microbiol. – 2000. – Vol. 42, No. 2. – P. 759-763.
2 Kamps B. S., Hoffmann C., Preiser W. Influenza Report 2006 / B. S. Kamps, С. Hoffmann, W. Preiser W. – Flying Publisher, 2006.
3 Wright S.M., Kawaoka Y., Sharp G.B., e.a. Interspecies transmission and reassortment of influenza Aviruses in pigs and turkeys in United States // Arm J Epidemiol. – 1992. – Vol. 136. – P. 448-97.
4 Blinov V.M., Kiselev O.I. An analyses of the potential areas of recombination in the hemmaglutinin genes of animal influenza viruses in relation to their adaptation to a new host-man // Vopr. Virusol. – 1993. – Vol.38, No 6. – P. 263-268.
5 Kida H., Ito T., Yasuda J., e.a. Potential for transmission of avian influenza viruses to pigs // J Gen Virol. – 1994. – Vol.74. – P.2183-2188.
6 Webster R.G. The importance of animal influenza for human disease // J. Vac. – 2002. – Vol. 20, No. 2. – P.16-20.
7 Hiromoto Y., Yamazaki Y., Fukushima T., e.a. Evolutionary characterization of the six internal genes of H5N1 human influenza A virus // J Gen Virol. – 2000. – Vol.81. – P. 1293-1303.
8 Flandorfer A., Garcia-Sastre A., Basler C. and Palese P. Chimeric influenza A viruses with a functional influenza B virus neuraminidase or hemagglutinin // J. Virol. – 2003. – Vol.77. – Р. 9116-9123.
9 Fouchier R. A. et al. Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene // J Clin Microbiol. – 2000. – Vol.38. – P. 4096-4101.
10 Boom R., Sol C., Salimans M. Rapid and simple method for purification of nucleic acids // J. Clin Microbiol. – 1990. – Vol. 28. – P. 495-503.
11 WHO (2002). WHO manual on animal influenza diagnosis and surveillance. Geneva, World Health Organization (document WHO/CDS/CSR/NCS/2002.5, available at: http://www.who.int/csr/resources/publications/influenza/en/whocdscsrncs20025rev.pdf
12 Патент «Способ бесфенольного выделения нативной РНК высокопатогенного вируса гриппа птиц из вируссодержащего материала для постановки полимеразной цепной реакции», № 60599 от 29.01.2007 Султанкулова К.Т., Сандыбаев Н.Т., Зайцев В.Л., Жолдыбаева Е.В., Строчков В.М., Мамадалиев С.М.
13 Spackman E. Avian influenza virus detection and quantitation by real-time RT-PCR // J. Methods Mol Biol. – 2014. – Vol. 1161. – P. 105-108.
14 Bin Zhou, Matthew E. Donnelly., Derek T. Scholes, Single-Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses // J. Virol. – 2009 Oct. – Vol. 83(19). – P. 10309-10313.
References
1 Quilivan M, Cullinane A, Nelly M, et al. (2000) Comparison of Sensitivities of Virus Isolation, Antigen Detection and Nucleic Acid Amplification for Detection of Equine Influenza Virus. Clin Microbiol, 42(2):759-763.
2 Kamps BS, Hoffmann C, Preiser W (2006) Influenza Report 2006 (Kamps BS, Hoffmann С, Preiser W), Flying Publisher
3 Wright SM, Kawaoka Y, Sharp GB, e.a. (1992) Interspecies transmission and reassortment of influenza Aviruses in pigs and turkeys in United States. Arm J Epidemiol, 136:448-97.
4 Blinov VM, Kiselev OI (1993) An analyses of the potential areas of recombination in the hemmaglutinin genes of animal influenza viruses in relation to their adaptation to a new host-man. Vopr Virusol, 38(6):263-268.
5 Kida H, Ito T, Yasuda J, e.a. (1994) Potential for transmission of avian influenza viruses to pigs. J Gen Virol, 74:2183-2188.
6 Webster RG (2002) The importance of animal influenza for human disease. J. Vaс, 20(2):16-20.
7 Hiromoto Y, Yamazaki Y, Fukushima T, e.a. (2000) Evolutionary characterization of the six internal genes of H5N1 human influenza A virus. J Gen Virol. 81:1293-1303.
8 Flandorfer A, Garcia-Sastre A, Basler CF and Palese P (2003) Chimeric influenza A viruses with a functional influenza B virus neuraminidase or hemagglutinin. J. Virol, 77:9116-9123.
9 Fouchier RA et al. (2000) Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene. J Clin Microbiol, 38:4096-4101.
10 Boom R, Sol C, Salimans M (1990) Rapid and simple method for purification of nucleic acids. J. Clin Microbiol, 28:495-503.
11 WHO (2002). WHO manual on animal influenza diagnosis and surveillance. Geneva, World Health Organization (document WHO/CDS/CSR/NCS/2002.5, available at: http:www.who.int/csr/resources/publications/influenza/en/whocdscsrncs20025rev.pdf
12 Patent «A method for the phenol-free isolation of native RNA of a highly pathogenic avian influenza virus from a virus-containing material for polymerase chain reaction» № 60599 from 29.01.2007 Sultankulova KT, Sandybayev NT, Zaitsev VL, Zholdybaeva EV, Strochkov VM, Mamadaliev SM (In Russian)
13 Spackman E (2014) Avian influenza virus detection and quantitation by real-time RT-PCR. J. Methods Mol Biol, 1161:105-108.
14 Bin Z, Matthew ED, Derek TS (2009) Single-Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses. J. Virol, 83(19):10309-10313.
2 Kamps B. S., Hoffmann C., Preiser W. Influenza Report 2006 / B. S. Kamps, С. Hoffmann, W. Preiser W. – Flying Publisher, 2006.
3 Wright S.M., Kawaoka Y., Sharp G.B., e.a. Interspecies transmission and reassortment of influenza Aviruses in pigs and turkeys in United States // Arm J Epidemiol. – 1992. – Vol. 136. – P. 448-97.
4 Blinov V.M., Kiselev O.I. An analyses of the potential areas of recombination in the hemmaglutinin genes of animal influenza viruses in relation to their adaptation to a new host-man // Vopr. Virusol. – 1993. – Vol.38, No 6. – P. 263-268.
5 Kida H., Ito T., Yasuda J., e.a. Potential for transmission of avian influenza viruses to pigs // J Gen Virol. – 1994. – Vol.74. – P.2183-2188.
6 Webster R.G. The importance of animal influenza for human disease // J. Vac. – 2002. – Vol. 20, No. 2. – P.16-20.
7 Hiromoto Y., Yamazaki Y., Fukushima T., e.a. Evolutionary characterization of the six internal genes of H5N1 human influenza A virus // J Gen Virol. – 2000. – Vol.81. – P. 1293-1303.
8 Flandorfer A., Garcia-Sastre A., Basler C. and Palese P. Chimeric influenza A viruses with a functional influenza B virus neuraminidase or hemagglutinin // J. Virol. – 2003. – Vol.77. – Р. 9116-9123.
9 Fouchier R. A. et al. Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene // J Clin Microbiol. – 2000. – Vol.38. – P. 4096-4101.
10 Boom R., Sol C., Salimans M. Rapid and simple method for purification of nucleic acids // J. Clin Microbiol. – 1990. – Vol. 28. – P. 495-503.
11 WHO (2002). WHO manual on animal influenza diagnosis and surveillance. Geneva, World Health Organization (document WHO/CDS/CSR/NCS/2002.5, available at: http://www.who.int/csr/resources/publications/influenza/en/whocdscsrncs20025rev.pdf
12 Патент «Способ бесфенольного выделения нативной РНК высокопатогенного вируса гриппа птиц из вируссодержащего материала для постановки полимеразной цепной реакции», № 60599 от 29.01.2007 Султанкулова К.Т., Сандыбаев Н.Т., Зайцев В.Л., Жолдыбаева Е.В., Строчков В.М., Мамадалиев С.М.
13 Spackman E. Avian influenza virus detection and quantitation by real-time RT-PCR // J. Methods Mol Biol. – 2014. – Vol. 1161. – P. 105-108.
14 Bin Zhou, Matthew E. Donnelly., Derek T. Scholes, Single-Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses // J. Virol. – 2009 Oct. – Vol. 83(19). – P. 10309-10313.
References
1 Quilivan M, Cullinane A, Nelly M, et al. (2000) Comparison of Sensitivities of Virus Isolation, Antigen Detection and Nucleic Acid Amplification for Detection of Equine Influenza Virus. Clin Microbiol, 42(2):759-763.
2 Kamps BS, Hoffmann C, Preiser W (2006) Influenza Report 2006 (Kamps BS, Hoffmann С, Preiser W), Flying Publisher
3 Wright SM, Kawaoka Y, Sharp GB, e.a. (1992) Interspecies transmission and reassortment of influenza Aviruses in pigs and turkeys in United States. Arm J Epidemiol, 136:448-97.
4 Blinov VM, Kiselev OI (1993) An analyses of the potential areas of recombination in the hemmaglutinin genes of animal influenza viruses in relation to their adaptation to a new host-man. Vopr Virusol, 38(6):263-268.
5 Kida H, Ito T, Yasuda J, e.a. (1994) Potential for transmission of avian influenza viruses to pigs. J Gen Virol, 74:2183-2188.
6 Webster RG (2002) The importance of animal influenza for human disease. J. Vaс, 20(2):16-20.
7 Hiromoto Y, Yamazaki Y, Fukushima T, e.a. (2000) Evolutionary characterization of the six internal genes of H5N1 human influenza A virus. J Gen Virol. 81:1293-1303.
8 Flandorfer A, Garcia-Sastre A, Basler CF and Palese P (2003) Chimeric influenza A viruses with a functional influenza B virus neuraminidase or hemagglutinin. J. Virol, 77:9116-9123.
9 Fouchier RA et al. (2000) Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene. J Clin Microbiol, 38:4096-4101.
10 Boom R, Sol C, Salimans M (1990) Rapid and simple method for purification of nucleic acids. J. Clin Microbiol, 28:495-503.
11 WHO (2002). WHO manual on animal influenza diagnosis and surveillance. Geneva, World Health Organization (document WHO/CDS/CSR/NCS/2002.5, available at: http:www.who.int/csr/resources/publications/influenza/en/whocdscsrncs20025rev.pdf
12 Patent «A method for the phenol-free isolation of native RNA of a highly pathogenic avian influenza virus from a virus-containing material for polymerase chain reaction» № 60599 from 29.01.2007 Sultankulova KT, Sandybayev NT, Zaitsev VL, Zholdybaeva EV, Strochkov VM, Mamadaliev SM (In Russian)
13 Spackman E (2014) Avian influenza virus detection and quantitation by real-time RT-PCR. J. Methods Mol Biol, 1161:105-108.
14 Bin Z, Matthew ED, Derek TS (2009) Single-Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses. J. Virol, 83(19):10309-10313.
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Akylbayeva, K. K., Shynybekova, G. O., Tlenchiyeva, T. M., Sultankulova К. Т., & Sandybaev, N. T. (2018). Лабораторная диагностика гриппа типов A и Б методом ПЦР. Вестник КазНУ. Серия биологическая, 70(1), 100–108. извлечено от https://bb.kaznu.kz/index.php/biology/article/view/1256
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