A new dual-targeting real-time RT-PCR assay for hepatitis D virus RNA detection

Yan Wang; Jeffrey S Glenn; Mark A Winters; Li-Ping Shen; Ingrid Choong; Ya-Lun Shi; Sheng-Li Bi; Li-Ying Ma; Hui Zeng; Fu-Jie Zhang

doi:10.1016/j.diagmicrobio.2018.05.016

A new dual-targeting real-time RT-PCR assay for hepatitis D virus RNA detection

Diagn Microbiol Infect Dis. 2018 Oct;92(2):112-117. doi: 10.1016/j.diagmicrobio.2018.05.016. Epub 2018 May 29.

Authors

Yan Wang¹, Jeffrey S Glenn², Mark A Winters², Li-Ping Shen³, Ingrid Choong⁴, Ya-Lun Shi⁵, Sheng-Li Bi³, Li-Ying Ma⁶, Hui Zeng⁷, Fu-Jie Zhang⁸

Affiliations

¹ State Key Laboratory of Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention, Beijing, P.R. China; Clinical and Research Center of Infectious Diseases, Beijing Ditan Hospital, Clinical Center for HIV/AIDS, Capital Medical University, Beijing, P.R. China.
² Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, Stanford, California, USA; Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA.
³ National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
⁴ Eiger BioPharmaceuticals, Inc. Palo Alto, California, USA.
⁵ Beijing Anapure BioScientific Co. Ltd, Beijing, China.
⁶ State Key Laboratory of Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention, Beijing, P.R. China.
⁷ Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China.
⁸ Clinical and Research Center of Infectious Diseases, Beijing Ditan Hospital, Clinical Center for HIV/AIDS, Capital Medical University, Beijing, P.R. China. Electronic address: [email protected].

PMID: 29941366
DOI: 10.1016/j.diagmicrobio.2018.05.016

Abstract

In this study, a real-time reverse transcription-polymerase chain reaction (real time RT-PCR) assay targeting 2 genetic segments was established to detect HDV RNA. Utilizing the World Health Organization International Standard for Hepatitis D Virus RNA, the lower limit of detection was 575 IU/mL, and the linearity of quantification ranged from 575,000 IU/mL to 575 IU/mL. 384 HBsAg-positive samples collected from China were tested by this method and HDV antibody detection. Eleven samples were positive for anti-HDV IgG which may persist after HDV resolution, 6 samples were HDV RNA positive, and 5 samples were positive for anti-HDV IgM. This assay showed more sensitivity than the detection of anti-HDV IgM. These data demonstrate that the real-time RT-PCR assay for HDV RNA could be implemented in the clinical detection of HDV infection in chronic HBV-infected patients in China.

Keywords: Detection; HDV; RNA; Real-time PCR.

MeSH terms

Adult
China
Female
Genotype
Hepatitis Antibodies / immunology*
Hepatitis D / diagnosis*
Hepatitis D / virology
Hepatitis Delta Virus / genetics
Hepatitis Delta Virus / immunology
Hepatitis Delta Virus / isolation & purification*
Humans
Male
RNA, Viral / genetics
Real-Time Polymerase Chain Reaction / methods
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Viral Load

Substances

Hepatitis Antibodies
RNA, Viral