Laboratory diagnosis of influenza types A and B by PCR

Abstract

This paper presents the method we developed for detecting RNA of influenza A and B viruses using multi-primer polymerase chain reaction. This technique is based on simultaneous laboratory diagnostics of influenza A and B viruses using polymerase chain reaction and differs by high specificity and sensitivity, which allows to detect minimum amount of influenza virus RNA copies in a test sample. During development of the given method we have selected such specific primers as InfAM63 and InfAM258 for influenza A virus and InfBM26, InfBM293 for influenza B virus. Specificity of the polymerase chain reaction was tested on several strains of the influenza A and B viruses, obtained from the collection of microorganisms of the RIBSP. The polymerase chain reaction identifies in a material the unique DNA fragments specific to influenza A and B viruses. The sensitivity of the test is determined by polymerase chain reaction with various dilutions of viral RNA. The sensitivity threshold of the influenza A and B viruses on the basis of polymerase chain reaction is 1х10² RNA copies (0.1 pg) in a sample. Diagnostics using polymerase chain reaction allows accurately and quickly (~ 5-6 hours) to detect the RNA of the influenza A and B viruses in a clinical material. It is possible to study the RNA of the influenza A and B viruses simultaneously.