微生物与感染
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微生物与感染  2020, Vol. 15 Issue (3): 158-165    DOI: 10.3969/j.issn.1673-6184.2020.03.004
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实时荧光核酸恒温扩增试验和定量反转录-聚合酶链反应对血清乙型肝炎病毒RNA定量检测的一致性评价
黄晨璐1,许伟1,胡乾坤1,张小楠1,李强1,黄玉仙1,2,陈良1
1.复旦大学附属公共卫生临床中心,上海,201508;2.复旦大学附属华山医院感染科,上海,200040
Evaluation of simultaneous amplification testing and quantitative reverse transcription-polymerase chain reaction for quantitative detecting serum hepatitis B virus RNA
HUANG Chenlu1, XU Wei1, HU Qiankun1, ZHANG Xiaonan1, LI Qiang1, HUANG Yuxian1,2, CHEN Liang1
1. Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China; 2. Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai 200040, China
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摘要: 为评价RNA实时荧光恒温扩增试验(simultaneous amplification testing,SAT)与定量反转录-聚合酶链反应(quantitative reverse transcription-polymerase chain reaction,qRT-PCR)检测血清样本中乙型肝炎病毒(hepatitis B virus,HBV) RNA的相关性与一致性,本研究收集了在复旦大学附属公共卫生临床中心就诊的212例患者的血清样本,其中慢性乙型肝炎(chronic hepatitis B,CHB)患者HBV DNA≥100 IU/mL 81例、HBV DNA<100 IU/mL 76例,以及作为对照的非HBV感染患者55例。采用SAT和qRT-PCR方法分别检测所有血清样本中的HBV RNA,对检测结果进行统计学分析。在HBV DNA≥100 IU/mL 的CHB患者中,SAT和qRT-PCR的HBV RNA阳性检出率均为95.06%(77/81)。2种方法具有较好的相关性与一致性(R2=0.803和CCC=0.882)。Bland Altman分析显示绝对偏倚为0.1 log copies/mL,相对偏倚为0.97%。配对t检验显示,结果无统计学差异(t =1.617,P =0.110)。在HBV DNA<100 IU/mL的CHB患者中,HBV RNA阳性检出率,SAT技术为98.68%(75/76), qRT-PCR方法为88.16%(67/76), 2种方法相关性与一致性较差(R2=0.326和CCC=0.438)。Bland Altman分析显示绝对偏倚为0.5 log copies/mL,相对偏倚为0.86%。配对t检验显示,结果有统计学差异(t =3.654,P<0.001)。在非HBV感染对照患者中,SAT技术和qRT-PCR方法均未检出HBV RNA阳性。结果提示,在HBV DNA≥100 IU/mL的CHB患者中应用SAT与qRT-PCR检测血清HBV RNA,其相关性与一致性较好,在HBV DNA<100 IU/mL 的CHB患者中相关性与一致性存在一定差异。
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黄晨璐1
许伟1
胡乾坤1
张小楠1
李强1
黄玉仙1
2
陈良1
关键词 乙型肝炎病毒核糖核酸定量反转录-聚合酶链反应实时荧光核酸恒温扩增试验    
Abstract:The purpose of the current study is to evaluate the correlation and consistency of simultaneous amplification testing (SAT) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for quantitative detecting serum hepatitis B virus RNA. 212 serum samples from Shanghai Public Health Clinical Center including 81 CHB patients with HBV DNA≥100 IU/mL, 76 CHB patients with HBV DNA 100 IU/mL and 55 patients without HBV infection were detected by SAT and qRT-PCR methods. In HBV DNA≥100 IU/mL CHB patients, 95.06% (77/81) samples were positive both in SAT and qRT-PCR method. SAT showed a relevantly good correlation and comparability with qRT-PCR (R2=0.803, CCC=0.882). Bland Altman analysis shows absolute bias was 0.1 log10 copies/mL and relative bias was 0.97%. The paired T test analysis of test results had no difference (t =1.617,P =0.110). In HBV DNA 100 IU/mL CHB patients, 98.68% (75/76) samples were positive in SAT method and 88.16% (67/76) samples were positive in qRT-PCR method. The statistical analysis of test results had difference (P<0.001). SAT showed a relevantly bad correlation and comparability with qRT-PCR (R2=0.326, CCC=0.438). Bland Altman analysis shows absolute bias was 0.5 log10 copies/mL and relative bias was 86%. The paired T test analysis of test results had difference (t =3.654,P<0.001). In 55 patients without HBV infection, all samples were negative in SAT method and qRT-PCR method. Correlation analysis shows strong correlation and consistency between SAT and qRT-PCR for CHB patients with HBV DNA≥100 IU/mL. Difference was found in the measurement of RNA using SAT and qRT-PCR for CHB patients with HBV RNA 100 IU/mL.
Key wordsHepatitis B virus    RNA    Quantitative reverse transcription-polymerase chain reaction    Simultaneous amplification testing
ZTFEL:  R373.2  
基金资助:上海市科委医学引导类(中、西医)科技支撑项目(17411969700),上海申康医院发展中心市级医院新兴前沿技术联合攻关项目(SHDC12015129)
通讯作者: 陈良   
引用本文:   
黄晨璐1,许伟1,胡乾坤1,张小楠1,李强1,黄玉仙1,2,陈良1. 实时荧光核酸恒温扩增试验和定量反转录-聚合酶链反应对血清乙型肝炎病毒RNA定量检测的一致性评价[J]. 微生物与感染, 2020, 15(3): 158-165.
HUANG Chenlu1, XU Wei1, HU Qiankun1, ZHANG Xiaonan1, LI Qiang1, HUANG Yuxian1,2, CHEN Liang1. Evaluation of simultaneous amplification testing and quantitative reverse transcription-polymerase chain reaction for quantitative detecting serum hepatitis B virus RNA. JOURNAL OF MICROBES AND INFECTIONS, 2020, 15(3): 158-165.
链接本文:  
http://jmi.fudan.edu.cn/CN/10.3969/j.issn.1673-6184.2020.03.004     或     http://jmi.fudan.edu.cn/CN/Y2020/V15/I3/158

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