High Resolution Melting Curve Analysis for Rapid Detection of Pyrazinamide Resistance in &amp;lt;i&amp;gt;Mycobacterium tuberculosis&amp;lt;/i&amp;gt; Clinical Isolates

Osman, Fatima; Ismail, Farzana; Osman, Ayman G.; Omar, Shaheed Vally; Said, Halima M.; Ismail, Nazir

doi:10.4236/jtr.2016.44019

Cited by 3 publications

(1 citation statement)

References 34 publications

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“…For example, Osman et al . reported a specificity of 63% for pncA mutations 38 , while, Juréen et al . reported a specificity of 97.3% in China 36 .…”

Section: Discussionmentioning

confidence: 96%

A Multinational Analysis of Mutations and Heterogeneity in PZase, RpsA, and PanD Associated with Pyrazinamide Resistance in M/XDR Mycobacterium tuberculosis

Ramirez-Busby

Rodwell

Fink

et al. 2017

Sci Rep

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Pyrazinamide (PZA) is an important first-line drug in all existing and new tuberculosis (TB) treatmentregimens. PZA-resistance in M. tuberculosis is increasing, especially among M/XDR cases. Noted issues with PZA Drug Susceptibility Testing (DST) have driven the search for alternative tests. This study provides a comprehensive assessment of PZA molecular diagnostics in M/XDR TB cases. A set of 296, mostly XDR, clinical M. tuberculosis isolates from four countries were subjected to DST for eight drugs, confirmatory Wayne's assay, and whole-genome sequencing. Three genes implicated in PZA resistance, pncA, rpsA, and panD were investigated. Assuming all non-synonymous mutations cause resistance, we report 90% sensitivity and 65% specificity for a pncA-based molecular test. The addition of rpsA and panD potentially provides 2% increase in sensitivity. Molecular heterogeneity in pncA was associated with resistance and should be evaluated as a diagnostic tool. Mutations near the N-terminus and C-terminus of PZase were associated with East-Asian and Euro-American lineages, respectively. Finally, Euro-American isolates are most likely to have a wild-type PZase and escape molecular detection. Overall, the 8-10% resistance without markers may point to alternative mechanisms of resistance. Confirmatory mutagenesis may improve the disconcertingly low specificity but reduce sensitivity since not all mutations may cause resistance. Pyrazinamide (PZA) is an important first-line drug recommended by the World Health Organization (WHO) for treatment of tuberculosis (TB)1 . Although most patients diagnosed with TB are successfully treated with first-line drugs, rifampicin (RIF), isoniazid (INH), ethambutol, and PZA, the incidence of multidrug resistant TB (MDR-TB) (resistant to first-line drugs) is increasing every year 1 . Along with this rate, the incidence of PZA resistance (PZA R ) is also increasing since over 60% of MDR-TB cases globally are also PZA R (rates vary regionally) 2 . In this manuscript we denote isolates that are resistant or susceptible to PZA by growth-based drug susceptibility testing (DST) as PZA R or PZA S , respectively. PZA is unique among anti-TB drugs in that, while it demonstrates powerful in vivo sterilizing activity, it exhibits no activity against actively growing Mycobacterium tuberculosis bacilli under normal culture conditions at neutral pH 3 . The drug appears to preferentially act against non-replicating persisters with low metabolic activity at acid pH in vitro or in vivo 4 . Importantly, most new drug regimens proposed for treatment of drug-resistant TB (DR-TB) show improved outcomes when combined with PZA 5-9 . Although growth-based PZA susceptibility testing is recommended by the WHO 10 , the method yields inconsistent results in some cases and suffers from a relatively high false resistance rate 11 .

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“…For example, Osman et al . reported a specificity of 63% for pncA mutations 38 , while, Juréen et al . reported a specificity of 97.3% in China 36 .…”

Section: Discussionmentioning

confidence: 96%

A Multinational Analysis of Mutations and Heterogeneity in PZase, RpsA, and PanD Associated with Pyrazinamide Resistance in M/XDR Mycobacterium tuberculosis

Ramirez-Busby

Rodwell

Fink

et al. 2017

Sci Rep

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Comprehensive review on mechanism of action, resistance and evolution of antimycobacterial drugs

Chauhan

Kumar

et al. 2021

Life Sciences

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High resolution melting assay as a reliable method for diagnosing drug-resistant TB cases: a systematic review and meta-analysis

Keikha

Karbalaei

2021

BMC Infect Dis

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Background Tuberculosis (TB) is one of the most contagious infectious diseases worldwide. Currently, drug-resistant Mycobacterium tuberculosis (Mtb) isolates are considered as one of the main challenges in the global TB control strategy. Rapid detection of resistant strains effectively reduces morbidity and mortality of world’s population. Although both culture and conventional antibiotic susceptibility testing are time-consuming, recent studies have shown that high resolution melting (HRM) assay can be used to determine the types of antibiotic resistance. In the present meta-analysis, we evaluated the discriminative power of HRM in detecting all drug-resistance cases of TB. Methods A systematic search was performed using databases such as Cochrane Library, Scopus, PubMed, Web of Science, and Google Scholar. Related studies on the effect of HRM in the diagnosis of drug-resistant (DR) TB cases were retrieved by April 2021. We used Meta-Disc software to evaluate the pooled diagnostic sensitivity and specificity of HRM for the detection of each type of drug-resistant cases. Finally, diagnostic value of HRM was characterized by summary receiver operating characteristic (SROC) curve and the area under the curve (AUC) method. Results Overall 47 studies (4,732 Mtb isolates) met our criteria and were included in the present meta-analysis. Sensitivity, specificity, and AUC of HRM were measured for antibiotics such as isoniazid (93%, 98%, 0.987), rifampin (94%, 97%, 0963), ethambutol (82%, 87%, 0.728), streptomycin (82%, 95%, 0.957), pyrazinamide (72%, 84%, 0.845), fluoroquinolones (86%, 99%, 0.997), MDR-TB (90%, 98%, 0.989), and pan-drug-resistant TB (89%, 95%, 0.973). Conclusions The HRM assay has high accuracy for the identification of drug-resistant TB, particularly firs-line anti-TB drugs. Therefore, this method is considered as an alternative option for the rapid diagnosis of DR-TB cases. However, due to heterogeneity of included studies, the results of HRM assays should be interpreted based on conventional drug susceptibility testing.

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High Resolution Melting Curve Analysis for Rapid Detection of Pyrazinamide Resistance in <i>Mycobacterium tuberculosis</i> Clinical Isolates

Cited by 3 publications

References 34 publications

A Multinational Analysis of Mutations and Heterogeneity in PZase, RpsA, and PanD Associated with Pyrazinamide Resistance in M/XDR Mycobacterium tuberculosis

A Multinational Analysis of Mutations and Heterogeneity in PZase, RpsA, and PanD Associated with Pyrazinamide Resistance in M/XDR Mycobacterium tuberculosis

Comprehensive review on mechanism of action, resistance and evolution of antimycobacterial drugs

High resolution melting assay as a reliable method for diagnosing drug-resistant TB cases: a systematic review and meta-analysis

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