부산대학교 도서관

Background: One of the major challenges in developing countries is lack of infrastructure in peripheral laboratories. The transportation of the sample for culture and / or NAAT based assays is difficult as it cannot be carried out at ambient temperature. Under these limiting factors, the microbes lose viability and hence culture results are false negative. Similarly, degradation of sample results in poor Nucleic Acid quality and extraction and hence the results are not reliable. Methods: We therefore tested whether it will be feasible to use dry samples in case of Multiplex PCR assays for Trichomonas. vaginalis, Chlamydia trachomatis and Neisseria gonorrhea. We collected samples in duplicate (n=133), one as dry swab and the other swab was immediately put in transport medium and brought to the laboratory at ambient temperature within 4-5 hours. Total gDNA was isolated from both the samples and used as template to check the performance of our in-house developed Multiplex PCR using dry and wet swabs swab samples (n=133). After confirmation of accuracy, we used further sample in the standardization of multiplex PCR using. Furthermore, we evaluated and compared performance of the multiplex PCR assays with dry and wet swabs. Results: Based on our conducted studies results, we observed high concordance of results irrespective of whether the total DNA was isolated from dry or wet swab. However, the intensity of the amplicon DNA band is lower when DNA is isolated from dry swab than that of gDNA isolated from wet swabs. This could be due to difference in extraction of total DNA from the two swabs. Out of 133 samples, 06 were positive for all the three infections using both dry and wet swabs while 127 were negative for all the three. In the onward Study, developed triplex PCR to identify these three pathogens simultaneously in a sample. All clinical samples were then tested using uniplex PCR for each pathogen and using triplex PCR for clinical evaluations. Conclusion: The clinical evaluation of Multiplex PCR showed as alternative method of accurate and confirmed diagnosis in low resources settings countries. [ABSTRACT FROM AUTHOR]