Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of &lt;em&gt;Mycobacterium tuberculosis&lt;/em&gt; by high-resolution melting analysis

Sirous, Mehrandokht; Khosravi, Azar Dokht; Tabandeh, Mohammad Reza; Salmanzadeh, Shokrollah; Ahmadkhosravi, Nazanin; Amini, Sirus

doi:10.2147/idr.s178831

Cited by 18 publications

(10 citation statements)

References 30 publications

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“…Mutation analysis of drug resistance genes of second-line drugs showed that only gyrA gene of OFX resistance mutated in four second-line anti-tuberculosis drugs, and the mutation sites were 74, 90, 91 and 94. This is consistent with the results of Wang Z, Sirous M and others [35][36] .From the above analysis of drug resistance gene site we could see that there were not only the genetic and molecular biological characteristics of the mainstream MDR-TB strains at home and abroad, but also has a few special characteristics. These characteristics will provide a new idea for rapid detection of MDR-TB in Urumqi in the future.…”

Section: Discussionsupporting

confidence: 91%

Gene Mutation Characteristics and Cluster Analysis of Multidrug-Resistant Mycobacterium Tuberculosis in Urumqi

Yang

Yadikaer²,

Chen

et al. 2021

Preprint

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Objective To understand the molecular biological characteristics of multidrug-resistant Mycobacterium tuberculosis in Urumqi by the analysis of gene mutation and clustering of multidrug-resistant Mycobacterium tuberculosis in Urumqi, to provid theoretical basis for the prevention and control of multidrug-resistant tuberculosis in the future. Methods The tuberculosis patients who were diagnosed, registered, cultured positive Mycobacterium tuberculosis and over 16 years old in Urumqi were studied. Geno Type MTBDR plus Assay and MIRU-VNTR were used to analysis the gene mutation and clustering of multidrug-resistant mycobacterium tuberculosis in Urumqi. Result Mutation gene included Isoniazid resistance gene katG 315; Rifampicin resistance gene rpoB 526, 531; Ethambutol resistance gene embB 206; Streptomycin resistance gene rpsL 43 and RRs 1401; Ofloxacin resistance gene gyrA 74, 90, 91 and 94. It was shown that the Beijing genotype was predominant (49.530%) among these319strains. There were 14 strains multi-drug resistant strains, it was predominant (73.684%) of multi-drug resistant strains. Multi-drug resistant prevalence of Beijing strains was 8.861%. The clustering rate of MDR-TB was 5.263% in Urumqi. Conclusion Molecular biological characteristics of multidrug-resistant Mycobacterium tuberculosis have some particularities in Urumqi. Therefore, modern molecular epidemiological techniques such as molecular biology and DNA fingerprinting can be used to improve the speed of diagnosis and to carry out case tracking and source investigation.

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Section: Discussionsupporting

confidence: 91%

Gene Mutation Characteristics and Cluster Analysis of Multidrug-Resistant Mycobacterium Tuberculosis in Urumqi

Yang

Yadikaer²,

Chen

et al. 2021

Preprint

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“…However, based on our results, other advantages of an HRMA assay and real-time PCR technology are its ability to detect non-viable, fastidious, and unculturable organisms that would otherwise be missed by culture. Sirous et al, 42 in other bacterias, showed that HRMA assay is a rapid, accurate, and cost-effective method possessing high sensitivity and specificity for the determination of antibiotic resistance. However, A PCR-positive, culture-negative specimen may reflect a real pathogen, yet detecting them would lead to a biased lower sensitivity and specificity value of the HRMA test.…”

Section: Figurementioning

confidence: 99%

<p>Distribution of Class B and Class A β-Lactamases in Clinical Strains of <em>Pseudomonas aeruginosa</em>: Comparison of Phenotypic Methods and High-Resolution Melting Analysis (HRMA) Assay</p>

Dehbashi¹,

Tahmasebi²,

Alikhani³

et al. 2020

IDR

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Background: There are various phenotypic methods for identifying class B and class A βlactamase enzymes in Pseudomonas aeruginosa. The purpose of this study was to compare the sensitivity and specificity of different phenotypic methods with HRMA assay to detect βlactamase-producing P. aeruginosa strains. Methods: Eighty-eight of P. aeruginosa isolates were collected from different specimens. Conventional double-disk test (DDT) and EDTA-imipenem microbiological (EIM) were performed to detect ESBL and MBL-producing strains, respectively. Meanwhile, the Modified Hodge test and Carba-NP test were performed on all carbapenem-resistant strains. HRMA method and sensitivity and specificity of primers were determined based on the melt curve temperature range. In all comparisons, PCR was considered as the gold standard. Results: Of the 402 isolates collected from different clinical specimens, 88 isolates of P. aeruginosa were identified. However, 43 strains were (48.88%) ESBL-producing, and 7 strains (7.95%) were MBL-producing. Also, using the Modified Hodge test and Carba-NP method, 11 (12.5%) and 19 (21.59%) strains were carbapenemase-producing, respectively. The results of the HRMA test revealed that genes coding for bla SHV , bla TEM , bla KPC , bla IMP , bla VIM, and bla GES were detected in 44.31%, 22.72%, 13.63%, 14.7%, 5.6%, and 2.27% of P. aeruginosa isolates. Nonetheless, for bla KPC and bla GES genes, sensitivity and specificity of the Carba-NP test were 90.47%, 94.87%, and 83.36%, 94.80%, respectively. However, sensitivity and specificity of MHT was 91.66%, 98.70%, and 77.77%, 96.42%, respectively. For bla SHV and bla TEM genes, sensitivity and specificity of DDT were 95.55%, 95.55%, and 86%, 83.50%, respectively. However, sensitivity and specificity of EMI were 77.77%, 97.59%, and 91.66%, 97.43% for bla VIM and bla IMP , respectively. Conclusion: The HRMA is a powerful, accurate, closed-tube, rapid method for detecting β-lactamase genes in P. aeruginosa. The high sensitivity and specificity of this method, along with phenotypic tests, play a useful role in increasing the predictive value of clinical reports.

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“…Out of the 509 potential studies, 52 studies met all inclusion criteria. (1, 7–59) A PRISMA flow diagram in Figure 1 illustrates the breakdown of number of articles excluded because of the specified criterion.…”

Section: Resultsmentioning

confidence: 99%

A Systematic Review of Mutations Associated with Isoniazid Resistance Points to Lower Diagnostic Sensitivity for Common Mutations and Increased Incidence of Uncommon Mutations in Clinical Strains of Mycobacterium tuberculosis

Valafar

2020

Preprint

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Molecular testing is rapidly becoming integral to the global tuberculosis (TB) control effort. Uncommon mechanisms of resistance can escape detection by these platforms and lead to the development of Multi-Drug Resistant (MDR) strains. This article is a systematic review of published articles that reported isoniazid (INH) resistance-conferring mutations between September-2013 and December-2019. The aims were to catalogue mutations associated with INH resistance, estimate their global prevalence and co-occurrence, and their utility in molecular diagnostics. The genes commonly associated with INH resistance, katG, inhA, fabG1, and the intergenic region oxyR-ahpC were considered in this review. In total, 52 articles were included describing 5,632 INHR clinical isolates from 31 countries. The three most frequently mutated loci continue to be katG315 (4,100), inhA-15 (786), and inhA-8 (105). However, the diagnostic value of inhA-8 is far lower than previously thought, only appearing in 25 (0.4%) INHR isolates that lacked a mutation at the first two loci. Importantly, of the four katG loci recommended by the previous systematic review for diagnostics, only katG315 was observed in our INHR isolates. This indicates continued evolution and regional differences in INH resistance. We have identified 58 loci (common to both systematic reviews) in three genomic regions as a reliable basis for molecular diagnostics. We also catalogue mutations at 49 new loci associated with INH resistance. Including all observed mutations provides a cumulative sensitivity of 85.1%. The most disconcerting is the remaining 14.9% of isolates that harbor an unknown mechanism of resistance, will escape molecular detection, and likely convert to MDR-TB, further complicating treatment. Integrating the information cataloged in this and other similar studies into current diagnostic tools is essential for combating the emergence of MDR-TB. Exclusion of this information will lead to an unnatural selection which will result in eradication of the common but propagation of the uncommon mechanisms of resistance, leading to ineffective global treatment policy and a need for region-specific regiments. Finally, the observance of many low-frequency resistance-conferring mutations point to an advantage of platforms that consider regions rather than specific loci for detection of resistance.

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Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of <em>Mycobacterium tuberculosis</em> by high-resolution melting analysis

Cited by 18 publications

References 30 publications

Gene Mutation Characteristics and Cluster Analysis of Multidrug-Resistant Mycobacterium Tuberculosis in Urumqi

Gene Mutation Characteristics and Cluster Analysis of Multidrug-Resistant Mycobacterium Tuberculosis in Urumqi

<p>Distribution of Class B and Class A β-Lactamases in Clinical Strains of <em>Pseudomonas aeruginosa</em>: Comparison of Phenotypic Methods and High-Resolution Melting Analysis (HRMA) Assay</p>

A Systematic Review of Mutations Associated with Isoniazid Resistance Points to Lower Diagnostic Sensitivity for Common Mutations and Increased Incidence of Uncommon Mutations in Clinical Strains of Mycobacterium tuberculosis

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