高敏与普通荧光定量PCR技术在不同病毒载量慢乙肝患者HBV-DNA检测的一致性及应用价值

Consistency and the value of high-sensitivity and conventional fluorescence quantitative PCR in different viral load in patients with chronic hepatitis B

目的比较高灵敏度与普通荧光定量聚合酶链式反应(qRT-PCR)法对乙型肝炎病毒DNA(HBV-DNA)定量检测结果的相关性与一致性。方法收集2020年8月-2021年6月中山大学附属第三医院感染性疾病科门诊患者的血浆标本511份。以Bland-Altman图分析2种检测方法间绝对与相对偏差,以组内相关系数(ICC)评价方法间一致性。高敏法HBV-DNA检出率与临床指标的关系用χ^(2)检验分析。结果在收集的511份慢性乙型肝炎(CHB)患者样本中,高灵敏度与普通荧光定量PCR法在<2×10^(3)、2×10^(3)~<2×10^(6)和2×10^(6)~IU/mL组的ICC分别为0.54、0.80和0.66,差异有统计学意义(P<0.05)。Bland-Altman法也显示类似的结果。对于HBV-DNA低于检测下限(普通PCR法)的样本,乙型肝炎表面抗原(HBsAg)≥1000 IU/mL、丙氨酸氨基转移酶(ALT)异常和未治疗的高灵敏度法检出率显著高于HBsAg<1000 IU/mL、ALT正常和抗病毒治疗(χ^(2)=6.67、5.00、30.51,P均<0.05)。结论普通荧光定量PCR法与高灵敏度法HBV-DNA结果存在差异,且在低病毒载量的样本中,两种方法的一致性较低。对于低水平病毒血症、普通法HBV-DNA检测低于检测下限、ALT异常、HBsAg≥1000 IU/mL、尚未进行抗病毒治疗的CHB患者,建议使用高灵敏度HBV-DNA进行检测。

Objective Compared the correlation and consistency of two assays including ordinary and high sensitivity for theHBV-DNA viral load measurement.Methods A total of 511 plasma samples were collected from outpatients in theInfectious Diseases Department of the Third Affiliated Hospital of Sun Yat-sen University from August 2020 to June 2021.Bland-Altman plots were performed for method comparison,and agreement was assessed with intraclass correlationcoefficient(ICC).The correlations between the HBV-DNA viral load and different clinical parameters were analyzed by theχ^(2)test.Results Of the 511 samples of chronic hepatitis B(CHB)patients,the high sensitivities with the commonfluorescence quantitative PCR method were 0.54,0.80,and 0.66 in the<2×10^(3),2×10^(3)-<2×10^(6)and 2×10^(6)-IU/m L group,respectively(P<0.05).Bland-Altman approach suggested similar results.For the HBV-DNA negative(conventional PCR)samples,the detectable rates(high-sensitivity PCR)were significantly higher in hepatitis B surface antigen(HBs Ag)≥1000 IU/m L,alanine aminotransferase(ALT)>upper limit of normal(ULN),and untreated compared with HBs Ag<1000IU/m L,ALT≤ULN,and treatment(χ^(2)=6.67,5.00,30.51,all P<0.05).Conclusions The high-sensitivity HBV-DNA fluorescence quantitative PCR was more sensitive at low viral loads than that of the conventional PCR.The CHB patients with HBV-DNA negative(conventional PCR),especially with abnormal ALT,HBe Ag-positive,HBs Ag≥1000 IU/m L,and treatment-na?ve require a high sensitivity test.