title: Detection of influenza A virus subtypes using a solid-phase PCR microplate chip assay.
abstract: A rapid and sensitive microplate chip based on solid PCR was developed to identify influenza A subtypes. A simple ultraviolet cross-linking method was used to immobilize DNA probes on pretreated microplates. Solid-phase PCR was proven to be a convenient method for influenza A screening. The sensitivity of the microplate chip was 10(-3) μg/mL for the enzymatic colorimetric method and 10(-4) μg/mL for the fluorescence method. The 10 sets of primers and probes for the microplate chip were highly specific and did not interfere with each other. These results suggest that the microplate chip based on solid PCR can be used to rapidly detect universal influenza A and its subtypes. This platform can also be used to detect other pathogenic microorganisms.
authors.affiliation: Basic Medical School of Zhengzhou University, Zhengzhou, China; College of Food and Biological Engineering, Zhengzhou University of Light Industry, Zhengzhou, China.
authors.forename: Xin-Cheng
authors.initials: XC
authors.lastname: Sun
authors.affiliation: Basic Medical School of Zhengzhou University, Zhengzhou, China; Henan Biotechnology Research Centre, Zhengzhou, China.
authors.forename: YunLong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Basic Medical School of Henan University of Traditional Chinese Medicine, Zhengzhou, China. Electronic address: bioylp66@126.com.
authors.forename: Liping
authors.initials: L
authors.lastname: Yang
authors.affiliation: Bioengineering Research Center of Henan Province, Zhengzhou, China.
authors.forename: HuiRu
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000818:Animals; D003124:Colorimetry / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D005470:Fluorometry / Q000379:methods; D006801:Humans; D009980:Influenza A virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes
keywords: Influenza A virus; Microplate chip; Solid-phase RT-PCR