Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis

Hu, Si-yu; Li, Guoli; Li, Hui; Liu, Xiaoli; Niu, Jianjun; Quan, Shengmao; Wang, Feng; Wen, Hao; Xu, Ye; Li, Qingge

doi:10.1128/jcm.03395-13

Cited by 38 publications

(33 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They can be addressed with molecular beacons, as they are able to distinguish sequences that differ by even a single nucleotide substitution (120)(121)(122). Next, they can be detected by identifying shifts in melting temperatures obtained by real-time PCR curve analysis (123)(124)(125)(126). SNaPshot analysis (Thermo Fisher Scientific) allows the detection of up to 10 SNPs in a single experiment with a capillary electrophoresis instrument (127).…”

Section: Methods Based On Nonrepetitive Sequencesmentioning

confidence: 99%

Methodological and Clinical Aspects of the Molecular Epidemiology of Mycobacterium tuberculosis and Other Mycobacteria

et al. 2016

View full text Add to dashboard Cite

SUMMARYMolecular typing has revolutionized epidemiological studies of infectious diseases, including those of a mycobacterial etiology. With the advent of fingerprinting techniques, many traditional concepts regarding transmission, infectivity, or pathogenicity of mycobacterial bacilli have been revisited, and their conventional interpretations have been challenged. Since the mid-1990s, when the first typing methods were introduced, a plethora of other modalities have been proposed. So-called molecular epidemiology has become an essential subdiscipline of modern mycobacteriology. It serves as a resource for understanding the key issues in the epidemiology of tuberculosis and other mycobacterial diseases. Among these issues are disclosing sources of infection, quantifying recent transmission, identifying transmission links, discerning reinfection from relapse, tracking the geographic distribution and clonal expansion of specific strains, and exploring the genetic mechanisms underlying specific phenotypic traits, including virulence, organ tropism, transmissibility, or drug resistance. Since genotyping continues to unravel the biology of mycobacteria, it offers enormous promise in the fight against and prevention of the diseases caused by these pathogens. In this review, molecular typing methods forMycobacterium tuberculosisand nontuberculous mycobacteria elaborated over the last 2 decades are summarized. The relevance of these methods to the epidemiological investigation, diagnosis, evolution, and control of mycobacterial diseases is discussed.

show abstract

Section: Methods Based On Nonrepetitive Sequencesmentioning

confidence: 99%

Methodological and Clinical Aspects of the Molecular Epidemiology of Mycobacterium tuberculosis and Other Mycobacteria

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Second, the MMCA assay could detect minor variant subpopulations with a sensitivity that was equal to that of COLD-PCR Sanger sequencing but better than that of PCR Sanger sequencing. The plasmid DNA experiments showed that both MMCA and COLD-PCR Sanger sequencing could reliably detect rtM204V when it was present at a prevalence of as low as 5% of the total (25). In this study, the SD values for T m were smaller than 1°C, while the change in T m (⌬T m ) values were larger than 4°C, ensuring the unambiguous detection of the mutations.…”

Section: Discussionmentioning

confidence: 78%

Detection of Anti-Hepatitis B Virus Drug Resistance Mutations Based on Multicolor Melting Curve Analysis

Mou

Athar

et al. 2016

J Clin Microbiol

Self Cite

View full text Add to dashboard Cite

Detection of anti-hepatitis B virus (HBV) drug resistance mutations is critical for therapeutic decisions for chronic hepatitis B virus infection.We describe a real-time PCR-based assay using multicolor melting curve analysis (MMCA) 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 5 copies per reaction and could detect a minor mutant population (5% of the total population) with the reverse transcriptase M204V amino acid mutation in the presence of the major wild-type population when the overall concentration was 10 4 copies/l. The assay could be finished within 3 h, and the cost of materials for each sample was less than $10. Clinical validation studies using three groups of samples from both nucleos(t)ide analog-treated and -untreated patients showed that the results for 99.3% (840/846) of the samples and 99.9% (8,454/8,460) of the amino acids were concordant with those of Sanger sequencing of the PCR amplicon from the HBV reverse transcriptase region (PCR Sanger sequencing). HBV DNA in six samples with mixed infections consisting of minor mutant subpopulations was undetected by the PCR Sanger sequencing method but was detected by MMCA, and the results were confirmed by coamplification at a lower denaturation temperature-PCR Sanger sequencing. Among the treated patients, 48.6% (103/212) harbored viruses that displayed lamivudine monoresistance, adefovir monoresistance, entecavir resistance, or lamivudine and adefovir resistance. Among the untreated patients, the Chinese group had more mutation-containing samples than did the Pakistani group (3.3% versus 0.56%). Because of its accuracy, rapidness, wide-range coverage, and cost-effectiveness, the real-time PCR assay could be a robust tool for the detection if anti-HBV drug resistance mutations in resource-limited countries. Hepatitis B is caused by the hepatitis B virus (HBV), an enveloped DNA virus that infects the liver, causing hepatocellular necrosis and inflammation (1). Chronic hepatitis B (CHB) infection affects approximately 248 million people worldwide and is a leading cause of liver-related morbidity and mortality, particularly in low-and middle-income countries (LMICs) (2). Patients with CHB can be successfully treated using nucleos(t)ide analogs (NAs), but drug-resistant HBV mutants frequently arise, leading to treatment failure and progression to liver disease (3). The development of drug resistance begins with mutations in the HBV polymerase gene, followed by an increase in the viral load and serum alanine aminotransferase levels several weeks to months later (4). Detection of drug resistance mutations is thus critical in prompt decision making for new therapeutic regimes (5, 6). A method enabling detection of NA resistance mutations should be reliable, rapid, and, in particular, easy to use and cost-effective when the aim is for it to be used in high-CHB-burden countries, which are often undeveloped a...

show abstract

“…Nevertheless, there are some commercial tests whose performance has not yet been assessed and published studies are required. A recently evaluated test is MeltPro TB/INH assay (Zeesan Biotech, China), aimed to detect INH resistance mutations in katG, inhA promoter, and ahpC promoter, with a clinical sensitivity and specificity of 90.8 % and 96.4 %, respectively, and 99.1 % concordance with DNA sequencing [71]. Genomic profile characterization by mass spectrometry has also been used to detect drug resistance.…”

Section: New Molecular Methods For Detecting Resistancementioning

confidence: 99%

Recent Advances in Tuberculosis Diagnosis: IGRAs and Molecular Biology

Molina-Moya

Latorre

Lacoma

et al. 2014

Curr Treat Options Infect Dis

View full text Add to dashboard Cite

Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Use of Real-Time-PCR-Based Melting Curve Analysis

Cited by 38 publications

References 32 publications

Methodological and Clinical Aspects of the Molecular Epidemiology of Mycobacterium tuberculosis and Other Mycobacteria

Methodological and Clinical Aspects of the Molecular Epidemiology of Mycobacterium tuberculosis and Other Mycobacteria

Detection of Anti-Hepatitis B Virus Drug Resistance Mutations Based on Multicolor Melting Curve Analysis

Recent Advances in Tuberculosis Diagnosis: IGRAs and Molecular Biology

Contact Info

Product

Resources

About