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Pathology 2018 Research & Reviews: Journal of Medical and Health Sciences
ISSN: 2319-9865 Page 71
October 08-09, 2018
Edinburgh, Scotland
17th International Conference on
Pathology & Cancer
Epidemiology
Chronic hepatitis B (CHB) caused by Hepatitis B virus infection
is a global public health issue. Individualized treatment of
CHB becomes increasingly important and a best individualized
treatment plan cannot be established without the information of
HBV genetic diagnosis. Nowadays, HBV DNA quantification by
real-time PCR is widely used in clinical diagnosis. However, there
is an urgent need to establish some simple and efficient clinical
detection methods for HBV drug resistance gene mutations
detection. To address these issues, we proposed a study on
the establishment of convenient and efficient methods for HBV
drug resistance gene mutations detection. We described a real-
time PCR-based assay using melting curve analysis that could
accurately detect 24 HBV nucleotide mutations at 10 amino
acid positions in the reverse transcriptase region of the HBV
polymerase gene. The two-reaction assay had a limit of detection
of five copies per reaction and could detect 5% rtM204V in the
presence of the wild-type when the overall concentration was 104
copies/μL. The assay could be finished within three h and the
material cost for each samplewas less than 10USD. Clinical study
using three groups of samples involving both nucleotide analogs-
treated and untreated patients showed that 99.3% (840/846)
samples and 99.9% (8454/8460) amino acids were concordant
with PCR sequencing. The six minor mutation containing
samples undetected by PCR sequencing were confirmed by co-
amplification at lower denaturation temperature-PCR sequencing.
In the treated patients, 48.6% (103/212) were mutant comprising
lamivudine-monoresistance, adefovir-monoresistance, entecavir
resistance and lamivudine+adefovir resistance, respectively.
Among the untreated patients, Chinese group had more mutation
containing samples than did the Pakistani group (3.3% vs 0.56%).
Because of its accuracy, rapidness, wide coverage and cost-
effectiveness, the real-time PCR assay could be a robust tool
for anti HBV drug resistance mutations detection in resources-
limiting countries.
Probe based melting curve analysis in anti-HBV drug resistance
gene mutation detection
Muhammad Ammar Athar1,2, Yi Mou1,2, Yuzhen Wu1,2, Ye Xu1,2, Jianhua Wu 3,
Zhenxing Xu3, Zulfiqar Hayder4, Saeed Khan5 , Muhammad Idrees Khan6, Muham-
mad Israr Nasir7, Yiqun Liao1 and Qingge Li1,2
1Xiamen University, China
2 Shenzhen Research Institute of Xiamen University, China
3Xiamen Hospital of Traditional Chinese Medicine, China
4Quaid-e-Azam medical College, Pakistan
5Dow University of Health Sciences, Pakistan
6University of the Punjab, Pakistan
7Liaquat National Hospital, Pakistan
RRJMHS 2018
Volume: 7