ABSTRACT
Epidemiological studies based on molecular methods for identification of Plasmodium vivax and Plasmodium falciparum is gaining importance, because the conventional methods like microscopy, cannot detect low-level parasitaemia and mixed infections. Even though different types of laboratory investigations for diagnosing malaria-like rapid antigen detection and QBC were developed, still there are some negative interpretations among convalescent cases identified in endemic areas. These issues can be overcome by using molecular techniques like multiplex PCR, nested PCR, real-time PCR and reverse transcriptase PCR. Among these methods real-time PCR has been shown to be more sensitive in studying the epidemiology of malaria. We wanted to standardize Multiplex PCR for the identification of Plasmodium species in a single reaction mix.METHODSA total of 52 blood samples were collected from suspected cases of clinical malaria which were tested for microscopy by using Leishman's stain and confirmed by conventional and multiplex PCR. Standardization of multiplex PCR for the identification of Plasmodium species in a single reaction mix was done for the diagnosis of malaria.RESULTSOut of 52 blood samples collected, about 38 (73.08 %) samples were confirmed with a multiplex PCR technique and only 34 (65.38 %) by microscopy. The four samples negative by microscopy were found to be Plasmodium falciparum. A significant correlation was found with the positive samples by conventional and standardized multiplex PCR.CONCLUSIONSMultiplex PCR is more useful for accurate diagnosis and epidemiological study for the detection of various species of the genus Plasmodium in a single-step reaction.