寡核苷酸芯片法、实时PCR 和测序法进行乙肝病毒 基因分型的比较

目的:比较寡核苷酸芯片法、实时荧光PCR和测序法在对慢性乙肝患者病毒基因分型的比较和方法学评价。方法:对126例不同基因型的慢性乙肝患者的血清样本分别用寡核苷酸芯片法、实时荧光PCR法和测序法进行基因分型,并评价各种方法的临床表现、所需时间和检测成本。结果:寡核苷酸芯片法、实时荧光PCR分别能检测到1%和0.1%比例的基因型。在126例慢性乙肝患者的临床样本中,寡核苷酸芯片法、实时荧光PCR和测序法分别检测出41(33%)、41(33%)和45(36%)例为B型,76(60%)、76(60%)、81(64%)例为C型。寡核苷酸芯片法、实时荧光PCR均检出9例B、C混合基因型。在三种检测方法中实时荧光PCR是最快速和廉价的。结论:寡核苷酸芯片法、实时荧光PCR能检出B、C混合基因型,而测序法只能检测出样本的主导基因型。

Comparison of Oligonucleotide Chip, Real-time PCR and Sequencing for Genotyping of Hepatitis B Virus

Objective: To compare the oligonucleotide chip,real-time PCR and sequencing for genotyping of hepatitis B virus in Chinese patients with chronic hepatitis B.Methods: Clinical serum samples with different hepatitis B virus genotype from 126 chronic hepatitis B patients were tested for hepatitis B virus genotypes by oligonucleotide chip,real-time PCR and sequencing of PCR products,respectively.Then,clinical performances,time required and costs of the three assays were evaluated.Results: Oligonucleotide chips and real-time PCR can detect 1 % and 0.1 % genotypes,respectively,in mixed samples.In the 126 clinical samples from patients with chronic hepatitis B,genotype B was detected in 41(33 %),41(33 %) and 45(36 %) samples,and genotype C in 76(60 %),76(60 %) and 81(64 %) samples,by oligonucleotide chip,real-time PCR and sequencing,respectively.Oligonucleotide chip and real-time PCR detected mixed genotypes B and C in 9 samples.Realtime PCR was the rapidest and cheapest method among the three assays.Conclusions: Oligonucleotide Chip and real-time PCR are able to detect mixed genotypes,while sequencing only detects the dominant genotype in clinical samples.