Comparison of a novel bilayered medium with the conventional media for cultivation of Mycobacterium tuberculosis.
ABSTRACT There is resurgence of tuberculosis in recent years in spite of availability of comprehensive multidrug therapy. Conventional culture media require a long time for the appearance of growth of Mycobacterium tuberculosis, while the other methods are expensive. Hence, a rapid low cost and safe bilayered medium was developed for early growth and sensitivity testing of M. tuberculosis and the results were compared with those on Lowenstein Jensen medium, Middlebrook 7H10 and Kirchner's liquid media.
A specially designed bilayered medium, consisting of a lower layer of Lowenstein Jensen medium without malachite green and a top layer of Middlebrook 7H 10 medium with added antibiotics and antifungal agents was prepared. Sputum from clinically suspected cases of tuberculosis, pleural fluid and pus samples were inoculated on the bilayered medium along with the inoculation on other conventional media after proper decontamination and concentration of the samples. Antibiotic sensitivity pattern was determined against a few rapidly growing control and test strains by disc diffusion technique and the results could be recorded by 3 to 7 days.
Statistically significant (P< 0.001) isolation rate was obtained on this bilayered medium when compared with the other three media, being 81.7 per cent growth by 7 days. Antibiotic sensitivity test could be recorded by 3 days in case of the rapidly growing strains on this medium, and by 7 days in case of M. tuberculosis strains.
Bilayered medium produced rapid growth earliest by 48 h, higher isolation rates were achieved as compared to the other conventional media and drug sensitivity testing could also be carried out successfully. Thus, the bilayered medium can be used for obtaining early culture report.
- [Show abstract] [Hide abstract]
ABSTRACT: Background & objectives: There has been an extensive invasion of tuberculosis at the global level by multidrug resistant as well as extensively drug resistant organisms. Attempts to recover the pathogen in pure culture have frequently failed since the specimens are often highly contaminated and also due to use of insufficient or over-active decontamination procedures. Hence in the present study different methods of decontamination were tested to evaluate their independent efficacies for culture of Mycobacterium tuberculosis. Methods: A total of 359 samples (241 sputum, 59 urine, 50 endometrium biopsy, 9 pus samples) from clinically suspected cases of tuberculosis were subjected to four different methods of decontamination followed by inoculation in Lowenstein-Jensen medium (LJM), and bilayered medium (BLM) and Kirchner's liquid medium (KLM) to determine the influence of differential decontamination processes. Sputum scanty and positive specimens were graded and each sample was subjected to decontamination by four different techniques. Results: Treatment of specimens with 4 per cent NaOH yielded minimum recovery of pure cultures, while use of 2 per cent NaOH produced higher number of contaminants compared to other methods of decontamination. Addition of N-acetyl L-cystein (NALC) coupled with 2 per cent NaOH to the samples for decontamination provided fairly reasonable recovery, but the highest number of M. tuberculosis cultures could be obtained when the specimens were treated with tri-sodium phosphate and benzalkonium (TSPB). Among the sputum positive cases recovery of growth of M. tuberculosis was higher with greater number of bacilli present in the specimens. Regarding the influence of culture media, BLM produced not only rapid growth, but reasonably higher rate of isolation of M. tuberculosis. Interpretation & conclusions: Although use of TSPB was found to be an efficient method of decontamination for successful isolation of M. tuberculosis from contaminated samples, both NALC+ 2 per cent NaOH and TSPB also showed significant recovery of M. tuberculosis cultures in BLM that can facilitate early diagnosis and initiation of treatment.The Indian Journal of Medical Research 10/2013; 138(4):541-8. · 1.66 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: Tuberculosis (TB) is the leading cause of death worldwide attributable to a single infectious disease agent. India has more new TB cases annually than any other country. In 2008, India accounted for a fifth of the estimated 9.4 million TB cases globally. There is an overwhelming need for improving TB diagnostics in India through the use of cost effective, patient-friendly methods appropriate to different tiers of the country health system. Substantial progress has been made in India in the field of TB diagnosis and serious efforts have been made to herald the development of diagnostic tests for pulmonary TB, extra pulmonary TB and MDR-TB. Diverse approaches have been attempted towards improving smear microscopy, rapid culture and for differentiation between the Mycobacterium tuberculosis complex and non-tuberculous mycobacteria. Several laboratories have developed in-house PCR assays for diagnosing TB with high accuracy. Approaches for distinguishing M. tuberculosis and/or Mycobacterium bovis infection and disseminated Mycobacterium avium complex infection in HIV-AIDS patients have also been described. Serological tests to detect antigens or antibodies to M. tuberculosis specific components by using cocktails of Excretory/Secretory protein antigens, Ag85 complex antigens, Hsp 65 antigen, RD1 antigens and Rapid Reverse Line Blot Hybridization assays to detect MDR-TB (mutations to rifampicin, isoniazid and streptomycin) have also been developed. Other methods like measurement of adenosine deaminase activity and use of luciferase reporter phages have also been explored for TB diagnosis. These advances in the Indian context are detailed in the present chapter. The validation and application of these methods in laboratory and public health settings is likely to result in improved TB diagnosis and contribute to effective disease management in India.Tuberculosis (Edinburgh, Scotland) 07/2011; 91(5):414-26. · 2.54 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: Tuberculosis globally results in almost 2 million human deaths annually, with 1 in 4 deaths from tuberculosis being human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS)-related. Primarily a pathogen of the respiratory system, aerobic Mycobacterium tuberculosis complex (MTBC) infects the lungs via the inhalation of infected aerosol droplets generated by people with pulmonary disease through coughing. This review focuses on M. tuberculosis transmission, epidemiology, detection methods and technologies.Infectious Diseases: Research and Treatment 01/2013; 6:39-50.