定量PCR-微流芯片法定量检测血清HBV DNA含量

Quantitative detection of HBV DNA in human serum by quantitative PCR-fluidic chip assay

目的应用定量PCR-微流芯片法定量检测临床血清标本HBV DNA含量,以了解该方法的灵敏度。方法将定量PCR-微流芯片法应用于10倍梯度稀释的HBV DNA参比品的定量检测,并与Taqman荧光定量法作比较;同时,应用这两种方法平行检测49份HBsAg阳性乙型肝炎患者的血清标本。结果Taqman荧光定量法可检测出104拷贝/mL的HBV DNA参比品,测得临床血清标本HBV DNA阳性率为63.27％(31/49),阳性血清的HBV DNA含量均值为9.60×106±19.54拷贝/mL;而PCR芯片法可测出103拷贝/mL HBV DNA参比品,测得临床血清标本HBV DNA的阳性率为77.55％(38/49),阳性血清的HBV DNA含量均值为6.95×106±15.43拷贝/mL。两者差异无显著性(P>0.05)。结论两种方法均能有效检测临床血清标本HBV DNA含量,而定量PCR一微流芯片法的敏感性更高,尤其适合于低病毒载量检测及患者抗病毒药物选择和疗效的监测。

Objective To explore the quantitative PCR-fluidic chip assay for quantitative detection of HBV DNA in clinical serum samples. Methods A 10-fold series dilution of standard HBV DNA was analyzed by quantitative PCR-fluidic chip, the serum HBV DNA quantification in 49 patients suffering from chronic hepatitis B with positive HBsAg was also detected. The results were compared with the detection of quantitative fluorescence PCR technique. Results The lowest detective value of standard HBV DNA was 104 copies/mL and serum HBV DNA positive rate was 63.27% (31/49), the average concentration of serum HBV DNA was 9. 60 x 106 + 19.54 copies/mL analyzed by the assay of quantitative fluorescence PCR; while the lowest detective value of standard HBV DNA was 10 copies/mL, serum HBV DNA positive rate was 77.55 % (38 /49) and the average concentration of HBV DNA was 6.95 x 10s + 15.43 copies/mL by quantitative PCR-fluidic chip assay. Significant difference was not found between the two methods ( P >0.05). Conclusions The two assays have the same important value in the clinical diagnosis, the quantitative PCR-fluidic chip assay has a higher sensitivity, and may be more suitable for evaluation of the therapeutic selection and efficacy for the patients with HBV infection.