Pyrosequencing for rapid detection of tuberculosis resistance to Rifampicin and Isoniazid in Syrian and Lebanese clinical isolates

Hamzé, Monzer; Ismail, Mohamad Bachar; Rahmo, Abdulkader; Dabboussi, Fouad

doi:10.1016/j.ijmyco.2015.05.007

Cited by 11 publications

(4 citation statements)

References 18 publications

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“…1 Multidrug-resistant tuberculosis is a form of TB infection caused by Mycobacterium tuberculosis (MTB) resistant to at least two of the most powerful first-line anti-TB drug treatments: Rifampicin and Isoniazid. 1,2 Xpert MTB/RIF assay is recommended by the World Health Organization (WHO) since 2010 as an initial test to diagnose MDR-TB. 3,4 Xpert MTB/RIF assay is a semi-quantitative nested real-time polymerase chain reaction (PCR) test that can detect deoxyribonucleic acid (DNA) of MTB rapidly.…”

Section: Introductionmentioning

confidence: 99%

“…6 However, the Xpert MTB/RIF assay only detects the rpoB gene mutations as Rifampicin resistance is a surrogate marker of MDR-TB. 2,5,7 Rifampicin-resistant TB (RR-TB) interpreted by the Xpert MTB/RIF assay diagnosed as MDR-TB was less accurate because the Xpert MTB/RIF assay cannot detect katG, inhA, and/or ahpC gene mutations that are associated with Isoniazid resistance. 8,9 Pyrosequencing is a DNA analysis based on nucleotide sequencing by inorganic pyrophosphate (PPi) synthesis to know nucleotide mutations directly and specifically.…”

Section: Introductionmentioning

confidence: 99%

“…inhA gene mutations, one katG, and inhA gene mutation, and 9 with no katG and inhA gene mutations as demonstrated in Table3.katG gene mutation detection based on the Pyrosequencing was found more than inhA gene mutation detection, with 27 subjects or 67.5% from 40 RR-TB subjects based on the Xpert MTB/ RIF assay. Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis study 2,10,11,13. Pyrosequencing interpreted one Isoniazid-resistant TB subject from 40 RR-TB subjects based on the Xpert MTB/RIF assay.…”

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confidence: 99%

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Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Choerunnisa

Rita

Tjandrawati

et al. 2020

mkb

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Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Choerunnisa

Rita

Tjandrawati

et al. 2020

mkb

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“…Molecular tests (e.g. polymerase chain reaction (PCR)) that target specific microorganisms have proven to be more rapid and sensitive than culturing [14], yet need a priori knowledge of the likely pathogenic species that could be present in a sample. As a complementary approach to culture, Sanger sequencing of the variable 16S rRNA gene has emerged for identification purposes [15].…”

Section: Introductionmentioning

confidence: 99%

Inguinal microbiome in patients undergoing an endovascular aneurysm repair: Application of next-generation sequencing of the 16S-23S rRNA regions

et al. 2019

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Background: Surgical site infection (SSI) remains a hazardous complication after vascular surgery. In this pilot study we investigated the inguinal microbiome in skin biopsies using histology and 16S-23S rDNA Next Generation Sequencing (NGS). Our hypothesis was that causative microorganisms of SSI are present in the inguinal microbiome. Methods: Data on surgical site infections and skin samples from the Percutaneous in Endovascular Repair versus Open (PiERO) trail were evaluated. Two patients with SSI were matched for age and comorbidity to eight matching patients of the PiERO trial. All patients were treated for an abdominal aortic aneurysm with endovascular repair. Nasal and perineal cultures were taken preoperatively to detect Staphylococcus aureus carriage. After disinfection with chlorhexidine, groin biopsies were taken to identify bacteria in deeper skin layers. All samples were subjected to histological analysis and culture-free 16S-23S rDNA NGS. Results: Staphylococcus aureus species were cultured in 5 out of 20 preoperative nasal and perineal swaps. Histology detected only a few bacteria. NGS of the 16S-23S rRNA regions identified DNA of bacterial species in all biopsies (20/20). Most identified genera and species proved to be known skin flora bacteria. No relation was found between SSIs and the preoperative microbiome. Conclusion:In this pilot study, an innovative analysis of the preoperative microbiome using 16S-23S rDNA NGS did not show a relation with the occurrence of a surgical site infection. No pathogenic bacterial species were present in the inguinal skin after disinfection with chlorhexidine.

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Evidence-Based Interventions for Antimicrobial Resistance in Conflict-Afflicted Arab Countries

Reffat¹,

Khoshnood²,

Dembry³

2021

Handbook of Healthcare in the Arab World

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Pyrosequencing for rapid detection of tuberculosis resistance to Rifampicin and Isoniazid in Syrian and Lebanese clinical isolates

Abstract: The high sensitivity and the short turnaround time combined with multilocus sequencing of several isolates in parallel make pyrosequencing an attractive method for drug resistance screening for MTB.

Cited by 11 publications

References 18 publications

Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Inguinal microbiome in patients undergoing an endovascular aneurysm repair: Application of next-generation sequencing of the 16S-23S rRNA regions

Evidence-Based Interventions for Antimicrobial Resistance in Conflict-Afflicted Arab Countries

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