Azher Kadhum Hassan and Vimala Gandhi S
mPCR is a sensitive assay and could be used as an accurate diagnostic method for detecting various types of microorganisms’ genome in low concentration in biological specimens. The demand for sensitive, rapid, safe and easy detection of PCR products has led researchers to a combination of this method with Rapid Card or ELISA. Conserved sequences were selected for design of semi nested primers. Samples were tested by RAPID Card and real- time PCR for detection of specific nucleic acid and viral genome respectively. Viral genome was extracted and reverse transcription was performed with M-Mulv and the cDNA kept at -80º C. The semi nested PCR products were analyzed by running on an agarose gel electrophoresis. Fifteen samples were tested with the Semi Nested PCR method. False positive or negative reactions were not observed. The results from other methods were compared with results obtained by electrophoresis. In gel electrophoresis, dilution of 1/10 was positive. Detection limits for gel electrophoresis as well as RAPID Card have been evaluated. It was shown that the Semi nested PCR method is more sensitive than conventional PCR.
