Comparison of mycobacterial identification methods study.docVIP

 PAGE \* MERGEFORMAT 8 Comparison of mycobacterial identification methods study Author: Liu Jinwei iron Kuang Pei Ning Ji Jin-Ping Song Zhongbin River Keywords:: Mycobacterium identification methods Comparison of micro-colony method, multiplex polymerase chain reaction (PCR) method and the traditional method used in clinical isolates of Mycobacterium strain identification results are reported as follows. Materials and Methods: Test strain: (1) standard strains: Mycobacterium tuberculosis H37RV, Mycobacterium bovis, Mycobacterium avium, Mycobacterium intracellular, Mycobacterium Kansas, M. scrofulaceum Mycobacterium fortuitum and the standard strains of Mycobacterium smegmatis , all purchased from Beijing Institute of Tuberculosis chest tumor. (2) clinical isolates: from my hospital inpatients TB sputum, pleural fluid and cerebrospinal fluid. Traditional typing qualification test: (1) culture identification test: p-nitrobenzoic acid by conventional preparation (PNB) and the thiophene -2 - carboxylic acid hydrazide (TCH) medium under test strain bacteria suspension (10-2/ml ) 0.1 ml inoculated, 37 ℃ culture, for 4 weeks to see results. (2) biochemical test: 68 ℃ heat resistant catalase test, nitrate reduction test and Tween 80 hydrolysis test, as a routine operation. Micro-colony observation: Take the test strains cultured at 37 ℃ for 2 ~ 3 weeks slant growths, with the preparation of sterile saline is equivalent to 10-3 mg wet bacteria / ml inoculated liquid, take 0.1 ml inoculated agar Kuangshi 3 , smooth evenly over the rear 36 ℃ with 5% CO2 the temperature boxes 3,7,12 d after training at 100 times the bacteria under a microscope observation of micro-morphology, record the results. Multi-PCR identification method: Multiple of three pairs of PCR primers are: MT1 and MT2, PT1 and PT2, IS5, and IS6, they were amplified 32 000 protein expression of the gene sequences [1], MTP40 protein gene sequences [2] and IS6110 insertion sequence [3]. DNA amplifi