Evaluation of PCR based DNA sequencing targeting mitochondrial larger subunit (mtLSU) region for the rapid detection of Pneumocystis jirovecii in sputum from HIV positive patients

Materials and methods A total of 104 sputum specimens from HIV positive and clinically suspected PJP patients attending Government Hospital of Thoracic Medicine during Dec 2009-Mar 2011 were included in the study. Direct staining of sputum specimens using KOH Calcoflour and Grocott methanamine silver staining (GMS) and nested PCR targeting mitochondrial larger subunit region (mtLSU) was optimized. The optimized PCR was applied on all clinical specimens and amplified products were subjected to DNA sequencing and the sequencing results were analyzed using BLAST analysis software to deduce the sequence homology.