HBV RNA检测在慢性乙型肝炎中的临床价值

doi:10.3969/j.issn.1673-8640.2022.01.015

检验医学 ›› 2022, Vol. 37 ›› Issue (1): 72-77.DOI: 10.3969/j.issn.1673-8640.2022.01.015

HBV RNA检测在慢性乙型肝炎中的临床价值

马艳婷¹, 苏曦¹, 黄斐¹, 王梦娴¹, 沈敏娜¹, 张春燕², 王蓓丽¹, 潘柏申¹, 郭玮¹()

1.复旦大学附属中山医院检验科,上海 200032
2.复旦大学附属中山医院厦门医院检验科,福建厦门 361015

收稿日期:2020-11-20 修回日期:2021-08-26 出版日期:2022-01-30 发布日期:2022-03-07
通讯作者: 郭玮
作者简介:郭玮,E-mail： guo.wei@zs-hospital.sh.cn。
马艳婷,女,1996年生,学士,技师,主要从事乙型肝炎病毒RNA临床应用研究。
苏曦,女,1977年生,主管技师,主要从事分子生物学检测工作。第一联系人：
马艳婷与苏曦对本研究具有同等贡献,并列为第一作者。
基金资助:
国家自然科学基金面上项目(81772263);国家自然科学基金面上项目(81972000);国家自然科学基金青年项目(81902139);国家重点研发计划“精准医学研究”重点专项课题(2017YFC0910003);上海市临床重点专科建设项目(shslczdzk03302);厦门市医疗卫生重点项目(YDZX20193502000002);复旦大学附属中山医院临床研究专项基金(2018ZSLC05)

Clinical role of HBV RNA detection in chronic hepatitis B

MA Yanting¹, SU Xi¹, HUANG Fei¹, WANG Mengxian¹, SHEN Minna¹, ZHANG Chunyan², WANG Beili¹, PAN Baishen¹, GUO Wei¹()

1. Department of Clinical Laboratory,Zhongshan Hospital,Fudan University,Shanghai 200032,China
2. Department of Clinical Laboratory,Xiamen Branch,Zhongshan Hospital,Fudan University,Xiamen 361015,Fujian,China

Received:2020-11-20 Revised:2021-08-26 Online:2022-01-30 Published:2022-03-07
Contact: GUO Wei

摘要/Abstract

摘要：

目的对检测乙型肝炎病毒（HBV）RNA的方法进行评估,比较不同乙型肝炎血清标志物模式患者血清HBV RNA水平及其与HBV DNA之间的相关性。方法对HBV RNA检测方法的精度密、灵敏度及线性范围进行评估。收集81例乙型肝炎表面抗原（HBsAg）阳性慢性乙型肝炎患者的血清,同时检测HBV RNA、HBV DNA及乙型肝炎血清学标志物。按乙型肝炎e抗原（HBeAg）是否阳性、不同乙型肝炎血清标志物模式及是否接受抗病毒药物治疗分别分组。结果 HBV RNA高值、低值样本的批内精密度分别为0.56%、2.30%,批间精密度分别为3.13%、6.03%,均符合临床要求;对接近厂商声明的检测下限的样本重复检测20次,检出率为100%,灵敏度符合要求;在HBV RNA 1.0×10²~1.0×10⁷拷贝/mL范围内成线性。HBeAg阳性组血清HBV RNA和HBV DNA载量均高于HBeAg阴性组（P<0.01、P<0.05）。在不同乙型肝炎血清标志物模式中,HBeAg阳性且乙型肝炎e抗体（HBeAb）阴性患者血清HBV RNA载量最高（P<0.01）。未治疗组血清HBV DNA及HBV RNA载量均高于治疗组（P<0.01、P<0.05）。在81例慢性乙型肝炎患者中,HBV RNA水平与HBV DNA载量、HBeAg阳性滴度呈正相关（r值分别为0.348、0.544,P<0.05）,与HBsAg滴度无相关性（r=0.04,P>0.05）。在HBeAg阳性的慢性乙型肝炎患者中,血清HBV RNA载量与血清HBV DNA载量呈正相关（r=0.338,P<0.05）。在HBeAg阴性的慢性乙型肝炎患者中,血清HBV RNA载量与血清HBV DNA载量无相关性（r=0.14,P>0.05）。结论血清HBV RNA是一种潜在的反映HBV活动的新型标志物,亦可用于HBV感染疗效监测。

关键词: 乙型肝炎病毒RNA, 乙型肝炎病毒DNA, 慢性乙型肝炎

Abstract:

Objective To evaluate the performance of a method for the detection of hepatitis B virus（HBV） RNA,to compare serum HBV RNA levels in patients with different results of serological markers of HBV infection,and to evaluate the relationship between HBV RNA and HBV DNA. Methods The performance of the method for HBV RNA detection was evaluated,which included precision,sensitivity and linearity. The sera of 81 patients with hepatitis B surface antigen（HBsAg）-positive chronic hepatitis B were collected,and the levels of HBV RNA,HBV DNA and serological markers were detected simultaneously. The patients were classified according to the different results of hepatitis B e antigen（HBeAg）,serological markers of HBV infection and medication. Results The intra-assay precisions of high and low HBV RNA values were 0.56% and 2.30%,and the inter-assay precisions were 3.13% and 6.03%,respectively,which all met the clinical requirements. The sensitivity of the assay was achieved by repeating the assay 20 times for samples close to the lower limit of detection stated by the manufacturer,and the detection rate was 100%,which met the clinical requirement. It was linear in the range of HBV RNA 1.0×10²-1.0×10⁷ copies/mL. Serum HBV RNA and HBV DNA loads in HBeAg-positive group were higher than those in HBeAg-negative group（P<0.01,P<0.05）,and the HBeAg-positive and hepatitis B e antibody（HBeAb）-negative group had the highest HBV RNA loads among all the groups with different results of serological markers of HBV infection（P<0.01）. Serum HBV DNA and HBV RNA loads were higher in untreated group than those in treated group（P<0.01,P<0.05）. In 81 patients with chronic hepatitis B,HBV RNA loads were positively correlated with HBV DNA load and HBeAg positive titer（r=0.348 and 0.544,respectively,P<0.05） and without HBsAg titer（r=0.04,P>0.05）. In HBeAg-positive patients with chronic hepatitis B,serum HBV RNA loads were positively correlated with serum HBV DNA load（r=0.338,P<0.05）. In HBeAg-negative patients with chronic hepatitis B,serum HBV RNA levels did not correlate with serum HBV DNA load（r=0.14,P>0.05）. Conclusions Serum HBV RNA level is a potential new marker for monitoring the viral activity and therapeutic efficacy of HBV infection.

Key words: Hepatitis B virus RNA, Hepatitis B virus DNA, Chronic hepatitis B

中图分类号:

R446.7

马艳婷, 苏曦, 黄斐, 王梦娴, 沈敏娜, 张春燕, 王蓓丽, 潘柏申, 郭玮. HBV RNA检测在慢性乙型肝炎中的临床价值[J]. 检验医学, 2022, 37(1): 72-77.

MA Yanting, SU Xi, HUANG Fei, WANG Mengxian, SHEN Minna, ZHANG Chunyan, WANG Beili, PAN Baishen, GUO Wei. Clinical role of HBV RNA detection in chronic hepatitis B[J]. Laboratory Medicine, 2022, 37(1): 72-77.

图/表 6

参考文献 20

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样本	批内精密度			批间精密度
样本	均值/（拷贝/mL）	标准差/（拷贝/mL）	CV/%	均值/（拷贝/mL）	标准差/（拷贝/mL）	CV/%
高值	2.51×10⁵	1.41×10³	0.56	5.62×10⁵	1.76×10⁴	3.13
低值	1.10×10³	2.53×10¹	2.30	1.41×10³	8.50×10¹	6.03

样本	批内精密度			批间精密度
样本	均值/（拷贝/mL）	标准差/（拷贝/mL）	CV/%	均值/（拷贝/mL）	标准差/（拷贝/mL）	CV/%
高值	2.51×10⁵	1.41×10³	0.56	5.62×10⁵	1.76×10⁴	3.13
低值	1.10×10³	2.53×10¹	2.30	1.41×10³	8.50×10¹	6.03

样本编号	稀释倍数	理论值/ （拷贝/mL）	实测值/ （拷贝/mL）
1	0	2.20×10⁷	2.19×10⁷
2	10	2.20×10⁶	2.72×10⁶
3	100	2.20×10⁵	2.15×10⁵
4	1 000	2.20×10⁴	1.85×10⁴
5	10 000	2.20×10³	2.03×10³
6	100 000	2.20×10²	1.89×10²

样本编号	稀释倍数	理论值/ （拷贝/mL）	实测值/ （拷贝/mL）
1	0	2.20×10⁷	2.19×10⁷
2	10	2.20×10⁶	2.72×10⁶
3	100	2.20×10⁵	2.15×10⁵
4	1 000	2.20×10⁴	1.85×10⁴
5	10 000	2.20×10³	2.03×10³
6	100 000	2.20×10²	1.89×10²

组别	例数	HBV RNA/（拷贝/mL）	HBV DNA/ （拷贝/mL）
HBeAg阳性组	40	1.2×10⁴（9.8×10²~3.0×10⁵）	8.0×10²（0~3.3×10⁵）
HBeAg阴性组	41	0 （0~2.7×10²）	0 （0~3.6×10³）
Z值		-5.866	-1.974
P值		0.000 1	0.048

HBV RNA检测在慢性乙型肝炎中的临床价值

Clinical role of HBV RNA detection in chronic hepatitis B

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参考文献 20

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Metrics

组别	例数	HBV RNA/ （拷贝/mL）	HBV DNA/ （拷贝/mL）
未治疗组	32	3.4×10³（0~1.5×10⁵）	1.3×10⁴（0~3.2×10⁶）
治疗组	49	1.3×10²（0~3.8×10³）	0（0~4.4×10²）
Z值		-2.025	-4.182
P值		0.043 0	0.000 1

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