On-site Detection Of Subtypes In Influenza A Virus

Influenza A virus belongs to the family Orthomyxoviridae,whose members are characterized by an eight-segmented RNA genome of negative polarity.The hemagglutinin(hemagglutinin,HA)and neuraminidase(neuraminidase,NA)are two important glycoprotein on surface of the gene segments.There are 18 subtypes of HA and 11 subtypes of NA based on antigenic differences in the surface glycoproteins.As present in 1878 of influenza outbreak,it has the highly potential to generate global pandemics.Systemic infection of animal and human beings,food health security and significant economic losses caused by the outbreak and epidemic of influenza A.Currently,the main diagnosis testing methods of A Influenza virus are immunofluorescence test and polymerase chain reaction.Due to the complex operation,they can not meet the requirements of rapid detection and on site diagnosis of influenza A virus.Loop-Mediated Isothermal Amplification(LAMP)is a rapidly developing new nucleic acid isothermal amplification technology that can amplify the target DNA or RNA rapidly,efficiently and specifically at constant temperature of 63 ?,which simplified the complexity of nucleic acid detection.Competitive annealing mediated isothermal amplification(CAMP)is an improved method by our laboratory which amplifies the nucleic acid in the constant temperature.There are similar advantages compared with LAMP.On the other hand,primer design of CAMP principle is simpler and the target DNA fragments are shorter.As a new generation of molecular biology detection methods,they are very suitable for the virus detection and diagnosis.In this study,the established LAMP and CAMP method was sucssessflully applied for the detection of pathogens such as H1-H18 and N1-N11 types of influenza a virus.Combination with microfluidic chip detection method,we realized the high flux and rapid detection for influenza a virus for rapid detection.The established methods have lots of advantges such as high sensitivity and specificity without nonspecific amplification of other types,which have a good application prospect.