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BACKGROUND 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. METHODS A 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. RESULTS Out 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. CONCLUSIONS Multiplex 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.

Introduction: A significant increase in hospital acquired urinary tract infections demand for improved diagnostic procedures, better clinical management, and prevention strategies. The present phenotypic and genotypic study was carried out from the urine samples which were collected from the patients presenting with urinary tract infection and with pre-existing urinary tract diseases and those without pre-existing urinary tract diseases for culture and sensitivity. Aim: The aim of the present study was to identify the bacteriological profile of isolates from patients with pre-existing urinary tract diseases and those without any pre-existing urinary tract diseases in a tertiary care hospital.Materials & Methods: This was a descriptive study, conducted at the department of Microbiology, Chettinad Hospital & Research Institute Kelambakkam. About 400 urine samples were collected from patients with clinical diagnosis of urinary tract infection, out of them 200 were from the patients with pre-existing urinary tract disease and 200 were from patients without pre-existing urinary tract disease. The samples were subjected to culture and drug sensitivity testing for identification of the pathogens and genotypic study for detecting genes coding for drug resistance.Result: The isolates in the group with preexisting urinary tract diseases were relatively resistant to almost all the panel of 1 st line and 2 nd line antibiotics than isolates obtained from those without pre-existing urinary tract diseases. The growth of Gram-positive cocci like Staphylococcus aureus and Gram-negative bacilli like Proteus species were seen to be isolated only in the patients with pre-existing urinary tract diseases and not in those without pre-existing urinary tract diseases. Genes like mec A from Staphylococcus aureus coding for methicillin resistance, Rh11 from Pseudomonas aeruginosa coding for biofilm formation, blaVIM & blaTEM from Escherichia coli, Klebsiella species, Pseudomonas aeruginosa were found in multidrug resistant isolates from patients with pre-existing urinary tract diseases. Conclusion: There was a clear association between the pre-existing urinary tract disease, the pathogen isolated and high degree antibiotic resistance. The high degree of resistance may be probably due to repeated use of antibiotics in these patients due to recurrent infections leading to subsequent resistance.

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In Southeast Asian countries, Chloroquine and Primaquine combination therapy is most successful in treating uncomplicated Plasmodium vivax infections, for preventing relapses.Later, resistance to Chloroquine has emerged resulting in therapeutic failures, and this encouraged the research to identify causative factors (1). Several molecular methods have been developed to identify the genetic mechanisms involved in resistance to chloroquine and other

Malaria is considered as a neglected tropical disease because of the therapeutic failures and lack of immunoprophylactic methods. Plasmodium species develop resistant genes by selective drug pressure, detection of such resistant parasitic strains by various molecular methods is significant in improving treatment modalities. Pvcrt-o, Pvmdr1 in Plasmodium vivax and Pfcrt-o in Plasmodium falciparum are the genes responsible for antimalarial resistance. In this study