000			02777nam a22002057a 4500
005			20161108094902.0
008			161108b2016 xxu||||| |||| 00| 0 eng d
041			_aeng
082			_a2612-T
100			_aFaiza Naz (2010-VA-288) _927044
110			_cProf. Dr. Masood Rabbani _926597
245			_aOptimization Of Multiplex Pcr For Simultaneous Detection Of Bacterial And Viral Water Borne Pathogens
260			_c2016.
300			_a68p.;
502			_aWaterborne illness is a serious issue throughout the world. Bacteria such as Salmonella spp., Shigella spp., E. coli spp. and from Viruses mostly Rotaviruses are involved in various waterborne outbreaks due to usage of contaminated water because of poor sanitation system mixing of waste material and fecal material with water, which can be transferred to human body by consuming such contaminated water Detection of these bacteria and virus from various foods by conventional method is not easy. Conventional methods are time consuming laborious and expensive. Now multiplex PCR is widely used for rapid detection of waterborne pathogens. The method is more sensitive and specific and can detect more than one pathogen in one single reaction mixture. This experimental design is developed to optimize the multiplex PCR reaction for detection of Salmonella spp., Shigella spp., Escherichia coli spp. and Rota virus. ATCC culture of Salmonella spp., Shigella spp., Escherichia coli spp., revived using standard culturing technique and multiplex PCR is optimized to amplify four different microbial genes simultaneously. A total 100 samples obtained from 10 towns, of Lahore. The samples were processed for multiplex PCR for detection of E. coli spp., Salmonella spp., Shigella spp. and Rota virus directly from water samples. With the amplification of 4 bacterial and viral genes simultaneously multiplex PCR was optimized. Water samples were obtained, to check the strength of planned experiment in the field. The samples were processed for and multiplex PCR for direct detection of Salmonella spp. and E. coli spp. directly from water samples. Similarly multiplex PCR was optimized with 3μl DNA template of each microbe , 56oC annealing temperature , 20pmol of every primer and 25μl of multiplex master mix. Multiplex PCR is more sensitive and specific. It is also time redeemable technique because conventional culturing method requires several days for the detection of waterborne pathogens but this technique wants expertise. This study was helpful to establish an optimized Multiplex PCR for the rapid and simultaneous detection of waterborne pathogens.
650			_aDepartment of Microbiology _94609
700			_aDr. Fareeha Akhtar _914923
700			_aDr. Sana Ullah Iqbal _924639
942			_cTH
999			_c9651 _d9650