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

BMC Infectious Diseases

Vanaja Kumar¹  Azger Dusthackeer¹  Gomathi Sivaramakrishnan¹  Balaji Subramanyam¹ 

[1] Department of Bacteriology, National Institute for Research in Tuberculosis (Indian Council of Medical Research), Chennai, India

关键词: Phage lysin;    Rapid Diagnosis;    M. tuberculosis;    DLRPA;   

Others  :  1158496
DOI  :  10.1186/1471-2334-13-44

 received in 2012-06-28, accepted in 2013-01-17,  发布年份 2013

【 摘 要 】

Background

Phage 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).

Methods

A 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.

Results

The 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%.

Conclusion

Phage lysin can be used to decontaminate sputum samples for the detection of M. tuberculosis by DLRPA directly from processed sputum specimens.

【 授权许可】

   
2013 Subramanyam et al.; licensee BioMed Central Ltd.

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