Laboratory Medicine ›› 2018, Vol. 33 ›› Issue (5): 437-441.DOI: 10.3969/j.issn.1673-8640.2018.05.015

• Orginal Article • Previous Articles     Next Articles

Flow fluorescent dot matrix technique for hepatitis B virus genotyping

ZHU Jun, ZHU Yuqing, ZHU Lingfeng, JIANG Lingli, XU Chong   

  1. Shanghai Center for Clinical Laboratory,Shanghai 200126,China
  • Received:2017-06-15 Online:2018-05-20 Published:2018-05-30

Abstract:

Objective To establish a flow fluorescent dot matrix technique (liquid microarray) for the determinations of hepatitis B virus (HBV) genotype B and C. Methods Specific primers against genotype B and C of HBV were designed to have 5' terminal labeled by biotin. After amplification by multiple polymerase chain reaction (PCR),the products were hybridized with different genotype specific oligonucleotide probes conjugated on the beads coded with different fluorescence. The beads were stained with streptavidin-phycoerythrin,and were determined by Luminex. A total of 67 samples were determined for genotyping by sequencing (24 cases of genotype B and 43 cases of genotype C). The efficiency of liquid microarray was evaluated. Results The results of liquid microarray were compared with those of sequencing,and the consistency rate was 94% (63/67). In 24 cases of genotype B determined by sequencing,only 1 case got double-positive signals from genotype B and C probes determined by liquid microarray. However,in 43 cases of genotype C,there were 3 cases getting double-positive signals. Conclusions Liquid microarray is accurate and effective for HBV genotyping,and it has the advance for the further determination of mixed infection of HBV with different genotypes.

Key words: Hepatitis B virus, Genotype, Multiple polymerase chain reaction, Hybridization, Flow fluorescent dot matrix technique

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