Carpanenamase-producing Enterobacteriaceae (CPE) has emerged as a threat to hospitalized patients. Phenotypic test such as Modified hodge test was less sensitive and specific especially to detect blaNDM-1 which is the most predominant genotype in this region. Nucleic acid amplification technology offers improved specificity and sensitivity. Failed amplification due to the presence of inhibitors is a limitation. In this study, we tried to use previous method described by Villumseen et al with some modification using another DNA extraction kit. Methods: Ten mls of sterile whole blood taken from nearly expired blood bag from blood bank was spiked with 200 μl of 0.5mcFarland bacterial suspension from thirty-six confirmed isolates of blaNDM-1 carbapenamase-producing Klebsiella pneumoniae in an aerobic Bactec Plus and incubated until the growth was detected. The blood specimen was subjected to DNA extraction method using Macherey-Nachel, Nucleospin® Blood QuickPure followed with multiplex PCR. Results: Out of the 36 isolates, 12 isolates revealed blaNDM-1 , 9 isolates revealed blaNDM-1 and blaOXA-48, 7 isolates revealed blaNDM-1, blaVIM and blaKPC genotypes that were amplified at cycle threshold of less than 30. Another 8 isolates could not pick up any genotypes possibly due to pipetting error as all the internal control were amplified. Eight true negative gram negative isolates underwent same procedure and none amplified at a cycle threshold less than 30. Conclusion: This modified method was proved to give a high yield of CPE genotypes with the cycle threshold was set at less than or equal to 30 and able to overcome the presence of PCR inhibitors.
Trigona thoracica propolis is known to have antimicrobial properties, however its antileptospiral properties and its synergistic effects with commonly prescribed antibiotics are scarcely documented. This study aimed to evaluate the antileptospiral properties of Trigona thoracica against pathogenic Leptospira species (spp.) and to study its synergistic effects with commonly prescribed antibiotics. Materials and Methods: The tested Leptospira serovars were Australis, Bataviae, Canicola and Javanica.Aqueous extract propolis (AEP) and ethanolic extracts propolis (EEP) were used. Broth dilution methods were used to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and the synergistic effects between the propolis and the tested antibiotics. The synergistic effects was evaluated by using the fractional inhibitory concentration (FIC) index. Morphological changes of the treated Leptospira were observed under a Scanning Electron Microscope (SEM). Results: The AEP and EEP were found to have antileptospiral properties against the tested Leptospiraspp. The synergy result showed that only combination of AEP and penicillin G against serovar Australis has demonstrated synergistic effect with the FIC index of 0.38. Morphological study using SEM showed significant structural changes of the treated Leptospira spp. Conclusions: The result suggests that Trigona thoracicapropolis could potentially be used as either a complimentary or an alternative therapeutic agent against pathogenic Leptospiraspp.