Phage lysin as a substitute for antibiotics to detect Mycobacterium tuberculosis from sputum samples with the BACTEC MGIT 960 system

Balaji, S.; Sivaramakrishnan, Gomathi; Dusthackeer, Azger; Selvakumar, N; Kumar, Vanaja

doi:10.1111/j.1469-0691.2011.03601.x

Cited by 13 publications

(7 citation statements)

References 16 publications

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“…Phage therapy was previously used therapeutically in humans (201) and to eliminate the contamination of food by food-borne bacterial pathogens (202). Bacteriophages were previously used by Subramanyam et al to decontaminate sputum samples instead of using antiseptics and/or antibiotics before performance of Mycobacterium species culture (203,204). The use of T1, T4, and phiX174-like E. coli phages was initially undertaken to decrease the number of E. coli colonies adhering to the urothelium and causing persistent urinary tract infections (205).…”

Section: Microbial Culturomicsmentioning

confidence: 99%

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

et al. 2015

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SUMMARY Bacterial culture was the first method used to describe the human microbiota, but this method is considered outdated by many researchers. Metagenomics studies have since been applied to clinical microbiology; however, a “dark matter” of prokaryotes, which corresponds to a hole in our knowledge and includes minority bacterial populations, is not elucidated by these studies. By replicating the natural environment, environmental microbiologists were the first to reduce the “great plate count anomaly,” which corresponds to the difference between microscopic and culture counts. The revolution in bacterial identification also allowed rapid progress. 16S rRNA bacterial identification allowed the accurate identification of new species. Mass spectrometry allowed the high-throughput identification of rare species and the detection of new species. By using these methods and by increasing the number of culture conditions, culturomics allowed the extension of the known human gut repertoire to levels equivalent to those of pyrosequencing. Finally, taxonogenomics strategies became an emerging method for describing new species, associating the genome sequence of the bacteria systematically. We provide a comprehensive review on these topics, demonstrating that both empirical and hypothesis-driven approaches will enable a rapid increase in the identification of the human prokaryote repertoire.

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Section: Microbial Culturomicsmentioning

confidence: 99%

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

et al. 2015

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“…The results indicated that phage lysin was comparable to antibiotic supplement in decontaminating sputum samples and in detecting M. tuberculosis [8]. The present work was aimed at evaluating phage lysin in the rapid detection of M. tuberculosis directly from the processed sputum samples using luciferase reporter phage (LRP) assay.…”

Section: Introductionmentioning

confidence: 99%

Phage lysin to control the overgrowth of normal flora in processed sputum samples for the rapid and sensitive detection of Mycobacterium tuberculosis by luciferase reporter phage assay

et al. 2013

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BackgroundPhage lysin, extracted from three bacteriophages was used in place of antibiotics to control the overgrowth of normal flora in processed sputum samples leading to the sensitive detection of Mycobacterium tuberculosis using diagnostic luciferase reporter phage assay (DLRPA).MethodsA total of 129 sputum samples were processed by modified Petroff’s method. Two Lowenstein Jensen slopes were inoculated from the processed sputum deposit thus obtained. The remaining deposits were transferred to 7 ml of Middlebrook 7H9 complete medium supplemented with phage lysin and incubated at 37°C. DLRPA was done using phAE129 at days 7, 9, 14 and 21. At the end of day 21, the samples were centrifuged and the pellets were inoculated on to 2 more LJ slopes to validate DLRPA results.ResultsThe sensitivity and specificity of DLRPA in detecting M. tuberculosis from sputum specimens was 90% and 81% respectively compared to conventional LJ culture. The agreement between the methods was 87%. The rate of contamination for DLRPA using phage lysin was 9.3%.ConclusionPhage lysin can be used to decontaminate sputum samples for the detection of M. tuberculosis by DLRPA directly from processed sputum specimens.

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“…As examples, ANC (nalidixic acid and colimycin) medium promotes the specific growth of Gram-positive bacteria, and Campylobacter blood agar is an enriched selective medium that contains 5 different antimicrobial agents. Antibiotics are used in Mycobacterium culture media primarily as a cocktail that includes polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin (27). Antiseptics, such as bromocresol purple, were used previously in agar culture media to select Enterobacteriaceae in particular, with different uses, such as the isolation of injured coliforms from drinking water (28).…”

Section: Selective Culture Mediamentioning

confidence: 99%

“…This noninvasive method obviates the requirement for gastric aspiration without a difference in pulmonary tuberculosis diagnosis (33). Finally, lytic phages have been used to decontaminate the normal flora from sputum before M. tuberculosis culture to replace the use of antibiotics (27,34).…”

Section: Sample Decontaminationmentioning

confidence: 99%

Current and Past Strategies for Bacterial Culture in Clinical Microbiology

et al. 2015

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SUMMARY A pure bacterial culture remains essential for the study of its virulence, its antibiotic susceptibility, and its genome sequence in order to facilitate the understanding and treatment of caused diseases. The first culture conditions empirically varied incubation time, nutrients, atmosphere, and temperature; culture was then gradually abandoned in favor of molecular methods. The rebirth of culture in clinical microbiology was prompted by microbiologists specializing in intracellular bacteria. The shell vial procedure allowed the culture of new species of Rickettsia . The design of axenic media for growing fastidious bacteria such as Tropheryma whipplei and Coxiella burnetii and the ability of amoebal coculture to discover new bacteria constituted major advances. Strong efforts associating optimized culture media, detection methods, and a microaerophilic atmosphere allowed a dramatic decrease of the time of Mycobacterium tuberculosis culture. The use of a new versatile medium allowed an extension of the repertoire of archaea. Finally, to optimize the culture of anaerobes in routine bacteriology laboratories, the addition of antioxidants in culture media under an aerobic atmosphere allowed the growth of strictly anaerobic species. Nevertheless, among usual bacterial pathogens, the development of axenic media for the culture of Treponema pallidum or Mycobacterium leprae remains an important challenge that the patience and innovations of cultivators will enable them to overcome.

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Phage lysin as a substitute for antibiotics to detect Mycobacterium tuberculosis from sputum samples with the BACTEC MGIT 960 system

Cited by 13 publications

References 16 publications

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

Phage lysin to control the overgrowth of normal flora in processed sputum samples for the rapid and sensitive detection of Mycobacterium tuberculosis by luciferase reporter phage assay

Current and Past Strategies for Bacterial Culture in Clinical Microbiology

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