Clearly, diagnosis tools that allow more rapid identification of MTB
and characterization of drug susceptibility patterns will greatly benefit the management buy MK-2206 of TB. Due to the long generation time of MTB, traditional method using solid media for Mycobacterium identification required 6–7 weeks for growth, species identification, and susceptibility testing. In the last decades use of DNA hybridization technologies and liquid radiometric culture systems, such as BACTEC 460 TB (Becton Dickinson diagnostic Instrument Systems, Sparks, Md) has significantly reduced time of identification of Mycobacterium and determination of the drug susceptibility patterns. 6, 7 and 8 The direct detection of MTB in clinical samples has further been accredited for use only with acid test bacillus smears positive sputum. In the method of Mycobacteriophage-based assay that could be detect several Mycobacterial species, including MTB and characterize drug susceptibility patterns within 24–48 h of obtained positive culture. This novel approach utilizes genetically engineered reporter phage to defect viable Mycobacteria, which upon LRP infection produces quantifiable luminescence. In the presence of drug resistant of bacilli, retain their viability undergo phage infection Dactolisib order and also produce luminescence. In this way, quantification of photons with a luminometer could be used to reveal susceptibility
profile of each isolates. In this study revealed that host range of phAE 129 demonstrating its ability to identify primary clinical isolated of M. tuberculosis and to develop new modified method using chitin for homogenizing and decontaminating sputum sample ideal for using on LRP assay. 9 and 10 The chitin is a mild decontaminating agent and it was dissolved to concentrate sulfuric acid and further diluted to 5% H2S04. The hydrolysis of chitin by acid produces Calpain acetic acid and chitosamine
which as mucolytic action against sputum process. 11 In the present study revealed that modified chitin H2S04 method of sputum processed LRP assay allows rapid and reliable recognition of organism in M. tuberculosis complex with high degrees of specificity and sensitivity. This diagnostic technology is a step closer to clinical readiness. The suspected 292 sputum samples were collected from identified pulmonary tuberculosis patients at various district level of Tamil Nadu, India. The samples were analyzed by standard procedure. These samples were collected individual container (Metconey bottles) recommended by standard laboratory procedure. The most commonly recommended containers are a sterile wide mouth jar with tightly fitted screw cap lid. The diagnostic specimens were collected before the initiation therapy. All specimens were transported to the laboratory and ideally processed at the earliest of the collection. Note: delay in process leads to falls negative culture and increased bacterial contamination.