A clinical, microbiological and economic analysis of a national service for the rapid molecular diagnosis of tuberculosis and rifampicin resistance in Mycobacterium tuberculosis

Drobniewski, Francis; Watterson, S A; Wilson, Stuart M.; Harris, G.

doi:10.1099/0022-1317-49-3-271

Cited by 46 publications

(41 citation statements)

References 29 publications

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“…18 The analysis of resistance to tuberculosis drugs in laboratory isolates using Mycobnet assesses the magnitude of the problem in the UK and indicates potential demographic risk factors. However, Mycobnet has limitations characteristic of any routine surveillance system such as incompleteness of information and reporting bias.…”

Section: Discussionmentioning

confidence: 99%

Antibiotic resistant tuberculosis in the United Kingdom: 1993-1999

Djuretic

Herbert²,

Drobniewski

et al. 2002

Thorax

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Section: Discussionmentioning

confidence: 99%

Antibiotic resistant tuberculosis in the United Kingdom: 1993-1999

Djuretic

Herbert²,

Drobniewski

et al. 2002

Thorax

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“…126 RMP resistance is important, since it is often a surrogate for MDR-TB. 127 About 95% of all M. TB RMP-resistant clinical isolates harbour specific mutations within a region of the rpoB gene. 128,129 In contrast to RMP, genotypic testing for INH resistance is much more complex and alterations in several genes including katG and inhA have been reported.…”

Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Dinnes¹,

Deeks²,

Kunst³

et al. 2007

Health Technol Assess

530

361

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“…The application of improved amplification techniques (in this case, nested PCR) allows greatly enhanced detection of smear-negative sputum specimens (8). Furthermore, rifampin resistance is detected using the amplification product as a template for sequencing for mutations conferring resistance, which is of practical interest as rifampin resistance represents a surrogate marker for multiple-drug resistant (MDR) M. tuberculosis (6,14). The rapid detection of tuberculosis and rifampin resistance can substantially reduce the cost of inappropriate isolation of patients with risk factors for MDR M. tuberculosis (6).…”

mentioning

confidence: 99%

Rapid Detection of Smear-Negative Mycobacterium tuberculosis by PCR and Sequencing for Rifampin Resistance with DNA Extracted Directly from Slides

2001

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Conventional methods for identification of Mycobacterium tuberculosis from culture can take 6 weeks. To facilitate the rapid detection of M. tuberculosis and to assess the risks of drug resistance, we developed a technique of eluting DNA directly from sputum slides and performing PCR for the detection of M. tuberculosis DNA, followed by sequencing the rpoB gene to detect rifampin resistance. This entire process requires only 48 h. Forty-seven sputum specimens submitted for microscopy for detection of acid-fast bacilli (AFB) and for mycobacterial culture and susceptibility testing were assessed after elution from the slides and extraction. M. tuberculosis-specific DNA was amplified in a nested PCR with previously described primers (primers rpo95-rpo293 and rpo105-rpo273), followed by analysis on a 4% agarose gel for a 168-bp product. Automated sequencing was performed, and the sequences were aligned against a database for detection of anomalies in the rpoB gene (codons 511 to 533) which indicate rifampin resistance. Of the 47 sputum specimens tested, 51% (24 of 47) were culture positive (time to positive culture, 2 to 6 weeks). Smears for AFB were positive for 58% (14 of 24) of the specimens and were negative for 42% (10 of 24) of the specimens. All 24 culture-positive sputum specimens (14 microscopy-positive and 10 microscopy-negative sputum specimens) were positive by PCR with eluates from the smears. Forty-nine percent (23 of 47) of the sputum specimens were negative for M. tuberculosis by smear, culture, and PCR. Of the isolates from the culture-positive samples, five were rifampin resistant by sequencing; all five were also rifampin resistant by in vitro susceptibility testing. Of these rifampinresistant M. tuberculosis isolates, two were microscopy negative for AFB. Patients who are negative for AFB and culture positive for M. tuberculosis can now be identified within a day, allowing institution of therapy and reducing isolation time and medical costs.In the last decade, tuberculosis has reemerged as one of the leading causes of death, killing nearly 3 million people annually. With an estimated 8.8 million new cases every year and the continued proliferation of drug resistance, the public health implications are immense (5, 14). Conventional methods for positive identification of Mycobacterium tuberculosis from culture can take up to 6 weeks, and it has been reported that tuberculosis can be transmitted from 17% of patients who are smear negative, culture positive for M. tuberculosis (1). To facilitate the rapid detection of M. tuberculosis we have developed a technique of eluting DNA directly from a sputum slide and performing a nested PCR amplification. Additionally, this PCR product (a section of the rpoB gene) may be sequenced to determine missense mutations, insertions, or deletions which correlate at Ͼ96% with rifampin resistance (13). This entire process requires only 48 h. MATERIALS AND METHODSForty-seven sputum specimens were obtained from the Microbiology Department at Specialty Laboratories for w...

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A clinical, microbiological and economic analysis of a national service for the rapid molecular diagnosis of tuberculosis and rifampicin resistance in Mycobacterium tuberculosis

Cited by 46 publications

References 29 publications

Antibiotic resistant tuberculosis in the United Kingdom: 1993-1999

Antibiotic resistant tuberculosis in the United Kingdom: 1993-1999

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Rapid Detection of Smear-Negative Mycobacterium tuberculosis by PCR and Sequencing for Rifampin Resistance with DNA Extracted Directly from Slides

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