Zastosowanie testu RT-nested-PCR do wykrywania materialu genetycznego wirusa grypy swin w materiale klinicznym

• Autorzy: Kowalczyk A , Markowska-Daniel I. , Pejsak Z.

Warianty tytułu

EN

Application of RT-nested-PCR for RNA detection of swine influenza virus in clinical samples

• Języki publikacji: PL

Abstrakty

EN

Pigs serve as major reservoirs of H1N1 and H3N2 influenza viruses which are endemic in pig populations worldwide and are responsible for one of the most prevalent respiratory diseases in pigs. Therefore the early detection and identification of such events are paramount in monitoring the spread of influenza viruses. Transfer of influenza A viruses from animal hosts to man may lead to the emergence of new human pandemic strains. The aim of the study was to prove the matrix gene useful for reverse transcription nested-PCR as sensitive and specific for the detection of swine influenza A viruses in clinical samples. 75 RNA-virus positive samples out of 235 samples, including nasal swabs, lung tissues and whole blood samples, were detected by RT-n-PCR. Using conventional virology method we isolated 33 SIV strains in embryonated chicken SPF eggs. The results of PCR were 100% in agreement with those of virus isolation. The limit of detection of SIV was 10-7 EID50/0.1 ml. These results demonstrate the usefulness of RT-n-PCR for the detection and identification of influenza A virus in clinical material.

Słowa kluczowe

PL

trzoda chlewna; swoistosc; metoda RT-nested PCR; wykrywanie wirusow; metody; wirus influenzy swin; czulosc

• Wydawca: -
• Czasopismo: Medycyna Weterynaryjna
• Rocznik: 2007
• Tom: 63
• Numer: 07
• Strony: 810-814
• Opis fizyczny: s.810-814,rys.,fot.,tab.,bibliogr.

Twórcy

autor

Kowalczyk A

• Panstwowy Instytut Weterynaryjny - Panstwowy Instytut Badawczy, Al.Partyzantow 57, 24-100 Pulawy

autor

Markowska-Daniel I.

autor

Pejsak Z.

Bibliografia

• 1.Aslanzadech J., Padilla B. B., Shanley J. D.: Evaluation of PCR and nested PCR for laboratory diagnosis of hepatitis C virus infection. Mol. Cell Probes. 1996, 10, 173-178.
• 2.Choi Y. K., Goyal S. M., Kang S. W., Farnham M. W., Joo H. S.: Detection and subtyping of swine influenza H1N1, H1N2 and H3N2 viruses in clinical samples using two multiplex RT-PCR assays. J. Virol. Methods 2002, 102, 53-59.
• 3.Di Trani L., Bedini B., Donatelli I., Campitelli L., Chiappini B., De Marco M. A., Delogu M., Buonavoglia C., Vaccari G.: A sensitive one-step real-time PCR for detection of avian influenza viruses using a MGB probe and an internal positive control. BMC Infect. Dis. 2006, 25, 87.
• 4.Ding L., Iimura E., Saijo K., Hamada H., Ohno T.: A speedy method to detect inserted DNA fragment in cell lines transfected with retroviral vectors. Cytotechnology 2000, 34, 243-252.
• 5.Easterday B. C., Van Reeth K.: Swine influenza, [w:] B. E. Straw, S. D'Allaire, W. L. Mengeling, D. J. Taylor (wyd.): Disease of swine. The Iowa State University Press, USA 1999, 277-290.
• 6.Ellis J. S., Zambon M. C.: Combined PCR-heteroduplex mobility assay for detection and differentiation of influenza A viruses from different animal species. J. Clin. Microbiol. 2001, 39, 4097-4102.
• 7.Fetzer C., Pesch S., Ohlinger V. F.: High risk of false positive results in a widely used diagnostic test for detection of the porcine reproductive and respiratory syndrome virus (PRRSV). Vet. Microbiol. 2006, 115, 21-31.
• 8.Ito T., Gorman O. T., Kawaoka Y., Bean W. J., Webster R. G.: Evolutionary analysis of the influenza A virus M gene with comparison of the M1 and M2 proteins. J. Virol. 1991, 65, 5491-5498.
• 9.Jóźwik A., Frymus T.: Comparison of the immunofluorescence assay with RT-PCR and nested PCR in the diagnosis of canine distemper. Vet. Res. Commun. 2005, 29, 347-359.
• 10.Kemp D. J., Churchill M. J., Smith D. B., Biggs B. A., Foote S. J., Peterson M. G., Samaras N., Deacon N. J., Doherty R.: Simplified colorimetric analysis of polymerase chain reactions: detection of HIV sequences in AIDS patients. Gene 1990, 94, 223-228.
• 11.Kho C. L., Mohd-Azmi M. L., Arshad S. S., Yusoff K.: Performance of an RT-nested PCR ELISA for detection of Newcastle disease virus. J. Virol. Methods 2000, 86, 171-183.
• 12.Markowska-Daniel I., Kowalczyk A.: Wpływ zmienności na międzygatunkową transmisję zakażeñ wirusem grypy. Medycyna Wet. 2006, 62, 981-987.
• 13.Pejsak Z., Markowska-Daniel I., Kowalczyk A., Jabłoñski A., Kozaczyñski W., Loda M.: Zaburzenia w rozrodzie związane z wybuchem grypy świñ w fermie wielkotowarowej. Medycyna Wet. 2005, 61, 1154-1159.
• 14.Stauber N., Brechtbuhl K., Bruckner L., Hofmann M. A.: Detection of Newcastle disease virus in poultry vaccines using the polymerase chain reaction and direct sequencing of amplified cDNA. Vaccine 1995, 13, 360-364.
• 15.Tao S. C., Jiang D., Lu H. L., Xing W. L., Zhou Y. X., Cheng J.: One-Tube Nested RT-PCR Enabled by Using a Plastic Film and its Application for the Rapid Detection of SARS-Virus. Biotechnol. Lett. 2004, 26, 179-183.
• 16.Wright K. E., Wilson G. A., Novosad D., Dimock C., Tan D., Weber J. M.: Typing and subtyping of influenza viruses in clinical samples by PCR. J. Clin. Microbiol. 1995, 33, 1180-1184.
• 17.Zhang W. D., Evans D. H.: Detection and identification of human influenza viruses by the polymerase chain reaction. J. Virol. Methods 1991, 33, 165-189.