title: Detection and typing of human-infecting influenza viruses in China by using a multiplex DNA biochip assay.
abstract: 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.
authors.affiliation: Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Infectious Diseases and Clinical Microbiology, Beijing Chao Yang Hospital, Capital Medical University, Beijing Institute of Respiratory Medicine, Beijing 100020, China.
authors.forename: Jiuxin
authors.initials: J
authors.lastname: Qu
authors.affiliation: Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Ba
authors.affiliation: Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jiuhong
authors.initials: J
authors.lastname: Dong
authors.affiliation: Translational Medicine Center, Guangdong Women and Children's Hospital, Guangzhou 511400, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Center for Systems Medicine, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China; Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China.
authors.forename: Aiping
authors.initials: A
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Dayan
authors.initials: D
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha, Hunan 410078, China.
authors.forename: Zanxian
authors.initials: Z
authors.lastname: Xia
authors.affiliation: College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Daxin
authors.initials: D
authors.lastname: Peng
authors.affiliation: State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Yuelong
authors.initials: Y
authors.lastname: Shu
authors.affiliation: Department of Infectious Diseases and Clinical Microbiology, Beijing Chao Yang Hospital, Capital Medical University, Beijing Institute of Respiratory Medicine, Beijing 100020, China; Lab of Clinical Microbiology and Infectious diseases, Centre of Respiratory and Critical Care Medicine, China Japan Friendship Hospital, Beijing 100029, China. Electronic address: caobin_ben@163.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Cao
authors.affiliation: Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; Center for Systems Medicine, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China; Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China. Electronic address: taijiao@moon.ibp.ac.cn.
authors.forename: Taijiao
authors.initials: T
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001717:Birds / Q000821:virology; D002681:China; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D053122:Influenza A Virus, H3N2 Subtype / Q000145:classification / Q000235:genetics; D053124:Influenza A Virus, H5N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D064766:Influenza A Virus, H7N9 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D058889:Molecular Typing / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D009976:Orthomyxoviridae Infections / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins
keywords: Biochip; Influenza virus; Multiplex RT-PCR; Typing