[RT-PCR for the determinationof the type of influenza virus circulating in the population].
Reverse transcription polymerase chain reaction (RT-PCR) was used to determine the type of flu virus that was circulating mostly in the population of the Basque Country in the 1999-2000 season. In this time period, 124 nasal and pharyngeal aspirations from persons with flu-like symptoms were analyzed. A parallel analysis was carried out using a conventional culture or a shell-vial in MDCK cell line, and immunofluorescence and multiple RT-PCR for the detection of influenza virus and its A and B types. Of the samples studied, 57 (45.96%) were positive for type A by the culture method and 64 (51. 61%) by RT-PCR of a genomic region that codifies the non-structural proteins NS1 and NS2. We also developed a rapid-detection method in which the transcription and amplification are carried out in a single step, using the same mix of transcription-amplification. Based on the results we can conclude that RT-PCR is a very sensitive method for the detection of the flu virus; we believe that its sensitivity, combined with its rapidity, makes it ideal for the detection of these respiratory viruses and the analysis of strains circulating in the population.
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Servicio de Microbiología Clínica, Hospital de Basurto, Facultad de Medicina, Universidad del País Vasco.
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D002460:Cell Line; D005455:Fluorescent Antibody Technique; D006801:Humans; D009980:Influenza A virus / Q000145:classification / Q000235:genetics; D009981:Influenza B virus / Q000145:classification / Q000235:genetics; D007251:Influenza, Human / Q000821:virology*; D001239:Inhalation; D009975:Orthomyxoviridae / Q000145:classification / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013030:Spain; D014764:Viral Proteins / Q000737:chemistry
D012367:RNA, Viral; D014764:Viral Proteins