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Identification of 54 Mycobacterial Species by PCR-Restriction Fragment Length Polymorphism Analysis of the hsp65 Gene (2001)
| Content Provider | CiteSeerX |
|---|---|
| Author | Tortoli, Enrico Fontana, Roberta |
| Abstract | A total of 121 reference and clinical strains of both slowly and rapidly growing mycobacteria belonging to 54 species were studied for restriction fragment length polymorphism of a PCR-amplified 439-bp segment of the gene encoding the 65-kDa heat shock protein. Restriction digests were separated by 10 % polyacrylamide gel electrophoresis (PAGE). By including a size standard in each sample, the restriction fragment profile was calculated using a computer-aided comparison program. An algorithm describing these 54 species (including 22 species not previously described) is proposed. We found that this assay based on 10 % PAGE provided a more precise estimate than that based on agarose gel electrophoresis of the real size of restriction fragments as deduced from the sequence analysis and allowed identification of mycobacteria whose PCR-restriction frag-ment length polymorphism analysis patterns were unequivocally identified by fragments shorter than 60 bp. Mycobacteria other than Mycobacterium tuberculosis (MOTT) are increasingly recognized as causing human infections (31). Conventional biochemical methods and phenotypic tests for species differentiation are laborious and time-consuming and frequently require specialized testing that is beyond the capac-ity of clinical laboratories. Genotypic methods for the identi- |
| File Format | |
| Publisher Date | 2001-01-01 |
| Access Restriction | Open |
| Subject Keyword | Mycobacterial Specie Pcr-restriction Fragment Length Polymorphism Analysis Hsp65 Gene Algorithm Describing Genotypic Method Conventional Biochemical Method 65-kda Heat Shock Protein Clinical Strain Restriction Fragment Profile Precise Estimate Restriction Digest Specie Differentiation Agarose Gel Electrophoresis Real Size Computer-aided Comparison Program Restriction Fragment Length Polymorphism Human Infection Clinical Laboratory Restriction Fragment Size Standard Specialized Testing Pcr-amplified 439-bp Segment Sequence Analysis Polyacrylamide Gel Electrophoresis Phenotypic Test |
| Content Type | Text |
