New universal primers for genotyping and resistance detection of low HBV DNA levels

Yongqing Tong, Bei Liu, Hui Liu, Hongyun Zheng, Jian Gu, Hang Liu, Min Lin, Yali Ding, Chunhua Song, Yan Li

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response. Asensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients' plasma is developed by PCR amplification of the DNA with novel universal primers. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients.

Original languageEnglish (US)
Article number232
JournalMedicine (United States)
Volume95
Issue number33
DOIs
StatePublished - Aug 1 2016

Fingerprint

Hepatitis B virus
DNA
Viral Drug Resistance
Mutation
Drug Resistance
Polymerase Chain Reaction
Nucleic Acid Databases
Viral Load
Therapeutics
Viruses

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Tong, Yongqing ; Liu, Bei ; Liu, Hui ; Zheng, Hongyun ; Gu, Jian ; Liu, Hang ; Lin, Min ; Ding, Yali ; Song, Chunhua ; Li, Yan. / New universal primers for genotyping and resistance detection of low HBV DNA levels. In: Medicine (United States). 2016 ; Vol. 95, No. 33.
@article{e585f29b52314e549f15f02dfab47fb7,
title = "New universal primers for genotyping and resistance detection of low HBV DNA levels",
abstract = "HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response. Asensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients' plasma is developed by PCR amplification of the DNA with novel universal primers. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients.",
author = "Yongqing Tong and Bei Liu and Hui Liu and Hongyun Zheng and Jian Gu and Hang Liu and Min Lin and Yali Ding and Chunhua Song and Yan Li",
year = "2016",
month = "8",
day = "1",
doi = "10.1097/MD.0000000000004618",
language = "English (US)",
volume = "95",
journal = "Medicine; analytical reviews of general medicine, neurology, psychiatry, dermatology, and pediatries",
issn = "0025-7974",
publisher = "Lippincott Williams and Wilkins",
number = "33",

}

Tong, Y, Liu, B, Liu, H, Zheng, H, Gu, J, Liu, H, Lin, M, Ding, Y, Song, C & Li, Y 2016, 'New universal primers for genotyping and resistance detection of low HBV DNA levels', Medicine (United States), vol. 95, no. 33, 232. https://doi.org/10.1097/MD.0000000000004618

New universal primers for genotyping and resistance detection of low HBV DNA levels. / Tong, Yongqing; Liu, Bei; Liu, Hui; Zheng, Hongyun; Gu, Jian; Liu, Hang; Lin, Min; Ding, Yali; Song, Chunhua; Li, Yan.

In: Medicine (United States), Vol. 95, No. 33, 232, 01.08.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - New universal primers for genotyping and resistance detection of low HBV DNA levels

AU - Tong, Yongqing

AU - Liu, Bei

AU - Liu, Hui

AU - Zheng, Hongyun

AU - Gu, Jian

AU - Liu, Hang

AU - Lin, Min

AU - Ding, Yali

AU - Song, Chunhua

AU - Li, Yan

PY - 2016/8/1

Y1 - 2016/8/1

N2 - HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response. Asensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients' plasma is developed by PCR amplification of the DNA with novel universal primers. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients.

AB - HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response. Asensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients' plasma is developed by PCR amplification of the DNA with novel universal primers. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients.

UR - http://www.scopus.com/inward/record.url?scp=84983554850&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84983554850&partnerID=8YFLogxK

U2 - 10.1097/MD.0000000000004618

DO - 10.1097/MD.0000000000004618

M3 - Article

C2 - 27537600

AN - SCOPUS:84983554850

VL - 95

JO - Medicine; analytical reviews of general medicine, neurology, psychiatry, dermatology, and pediatries

JF - Medicine; analytical reviews of general medicine, neurology, psychiatry, dermatology, and pediatries

SN - 0025-7974

IS - 33

M1 - 232

ER -