DEVELOPMENT OF A PCR BASED ASSAY FOR THE DETECTION OF MYCOPLASMA PNEUMONIAE

Abstract

In the present study we collected seven blood samples from patients infected with M. pneumonia.Genomic DNA was isolated from these blood samples and specific primers were designed for p1 gene coding for cytadhesin protein P1 of the M. pneumonia. The amplified product was cloned into pTZ57R/T cloning vector and transformed into E. coli strain DH5α. Plasmid was isolated from the transformed cells, digested and checked for gene product release. Also, colony PCR was used to confirm the cloning and transformation. The released gene product was eluted and sequenced.  The obtained sequence was 99% matching with a part of p1 gene available in the public nucleotide date base. In conclusion, this report describes a genomics-based PCR that provided rapid detection of M. pneumoniae. This PCR assay was developed for testing blood specimens suspected to harbor M. pneumoniae and is highly sensitive.