Detection and typing of human-infecting influenza viruses in China by using a multiplex DNA biochip assay.
Rapid identification of the infections of specific subtypes of influenza viruses is critical for patient treatment and pandemic control. Here we report the application of multiplex reverse transcription polymerase chain reaction (RT-PCR) coupled with membrane-based DNA biochip to the detection and discrimination of the type (A and B) and subtype (human H1N1, human H3N2, avian H5N1 and avian H7N9) of influenza viruses in circulation in China. A multiplex one-step RT-PCR assay was designed to simultaneously amplify the HA and NA genes of the four subtypes of influenza A viruses and NS genes to discriminate type A and B viruses. PCR products were analyzed by a membrane-based biochip. The analytical sensitivity of the assay was determined at a range of 2-100 copies/reactions for each of the gene transcripts. Eighty one clinical samples, containing 66 positive samples with evident seasonal influenza virus infections, were tested, which gives the clinical sensitivity and specificity of 95.5% and 100% respectively. For the avian influenza samples, 3 out of 4 H5N1 samples and 2 out of 2 H7N9 avian samples were correctly identified. We argue this method could allow a rapid, reliable and inexpensive detection and differentiation of human-infecting influenza viruses.
['Wang|Yongqiang|Y|', 'Qu|Jiuxin|J|', 'Ba|Qi|Q|', 'Dong|Jiuhong|J|', 'Zhang|Liang|L|', 'Zhang|Hong|H|', 'Wu|Aiping|A|', 'Wang|Dayan|D|', 'Xia|Zanxian|Z|', 'Peng|Daxin|D|', 'Shu|Yuelong|Y|', 'Cao|Bin|B|', 'Jiang|Taijiao|T|']
D000818:Animals D001717:Birds D002681:China D006801:Humans D053118:Influenza A Virus, H1N1 Subtype D053122:Influenza A Virus, H3N2 Subtype D053124:Influenza A Virus, H5N1 Subtype D064766:Influenza A Virus, H7N9 Subtype D005585:Influenza in Birds D007251:Influenza, Human D058889:Molecular Typing D060885:Multiplex Polymerase Chain Reaction D020411:Oligonucleotide Array Sequence Analysis D009975:Orthomyxoviridae D009976:Orthomyxoviridae Infections D012367:RNA, Viral D012680:Sensitivity and Specificity D014764:Viral Proteins
Typing Biochip Influenza virus Multiplex RT-PCR