Loading…
Development and Validation of Multiplex Quantitative PCR Assay for Detection of Helicobacter pylori and Mutations Conferring Resistance to Clarithromycin and Levofloxacin in Gastric Biopsy
More than half of the world's population is infected with , which can cause chronic gastritis. WHO has regarded clarithromycin-resistant as a high priority pathogen. Hence, accurate diagnosis and detection of clarithromycin- and levofloxacin-resistant strains is essential for proper management...
Saved in:
Published in: | Infection and drug resistance 2021-01, Vol.14, p.4129-4145 |
---|---|
Main Authors: | , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Summary: | More than half of the world's population is infected with
, which can cause chronic gastritis. WHO has regarded clarithromycin-resistant
as a high priority pathogen. Hence, accurate diagnosis and detection of clarithromycin- and levofloxacin-resistant
strains is essential for proper management of infection. The objective of this study was to develop and optimize multiplex quantitative PCR assay for detection of mutations associated with clarithromycin and levofloxacin resistance in
directly from the gastric biopsies.
Specific primers and probes were designed to amplify
and mutations in 23S rRNA and
genes. Singleplex and triplex qPCR assays were optimized and the assay's sensitivities and specificities were determined. The optimized multiplex qPCR assay was performed on 571 gastric biopsies.
In this study, 14.7% (84/571) of the gastric biopsies were positive for
by conventional methods and 23.8% (136/571) were positive by the
-qPCR with 96.4% sensitivity and 88.5% specificity, while the +LR and -LR were 8.72 and 0.04, respectively. The
-positive samples (n=136) were subjected to multiplex qPCR which detected A2142G and A2143G mutations in the 23S rRNA gene (20.6%, 28/136) conferring clarithromycin resistance and
mutations N87K, N87I, D91N, and D91Y (11.8%, 16/136) leading to levofloxacin resistance. The sensitivity and specificity of qPCR of 23S rRNA gene were 100% and 98.7%, respectively, while 100% and 99.8% for qPCR of
, respectively.
The effectiveness of this qPCR is that it is sensitive in detecting low bacterial load and will help in timely detection of clarithromycin- and levofloxacin-resistant strains, especially in case of mixed infections. Since it is culture independent, it can inform clinicians about antibiotics to be included in the first-line therapy, thereby improving the management of
infection at a much greater pace. |
---|---|
ISSN: | 1178-6973 1178-6973 |
DOI: | 10.2147/idr.s325056 |