Multiplex PCR for direct identification of Campylobacter Species in Human Stool
Journal Title: Journal of Biomedical and Pharmaceutical Research - Year 2015, Vol 4, Issue 3
Abstract
Background Campylobacter spp. are a major cause of bacterial gastroenteritis worldwide. Is one of the most frequently isolated bacteria from the feces of infants and children in developing countries, Conventional diagnostic methods for the detection and differentiation of Campylobacter species are tedious and time consuming, we have developed a multiplex PCR protocol using a novel combination of species-specific and virulence genes, The multiplex PCR protocol was capable of detecting the type strains and clinical isolates from all five species with a high degree of specificity. Methods: Molecular testing for Campylobacter Universal gene was done on 50 Stool specimens using Multiplex PCR technique. Stool specimens were collected from patients suffering from gastroenteritis, and kept at -20 OC till used .DNA Extraction was done by using Vivantis GF-1 Nucleic acid extraction kit (Vivantis, MALAYSIA). The amplification reaction was carried out in thermo cycler with program system consisting of ''( Initial denaturation step at 95°C for 6 min followed by 30 cycles of amplification (denaturation at 95°C for 0.5 min, annealing at 59°C for 0.5 min, and extension at 72°C for 0.5 min), ending with a final extension at 72°C for 7 min)'' PCR products were separated in a 1.5% agarose gel, then stained with ethidium bromide and viewed under gel documentation system. A result was considered positive when a band of the appropriate size was visible in the gel. Standard procedures for reducing contamination were strictly followed. Results: eight samples (16%) out of 50 were positive by Multiplex PCR, while 42 samples (84%) were negative. Conclusion: The multiplex PCR is very useful for direct detection and diagnosis of campylobacteriosis..
Authors and Affiliations
Khalid A Enan| Central Laboratory- Ministry of Science and Communication, Abdel Rahim M. El Hussein| Central Laboratory Ministry of Science and Communication, Elbrissi Atif| Department of Bacteriology, Central Laboratory- Ministry of Science and Communication, Mustafa E Yassin| M.Sc. Microbiology Department of Faculty of Medical Laboratory Sciences, Al Neelain University, Abbas Bakhit Mohammed Rahama| M.Sc. Microbiology Department of Faculty of Medical Laboratory Sciences, Al Neelain University, Isam M Elkhidir| Department of Microbiology and Parasitology, Faculty of Medicine, University of Khartoum, Khartoum, Sudan)
Keywords
Related Articles
- EP ID EP3560
- DOI -
- Views 369
- Downloads 24
TOXICITY OF KHAYA GRANDIFOLIOLA (AFRICAN MAHOGANY) BARK WATER EXTRACT AGANIST BIOMOPHLARIA SNAILS
Background: Molluscicides are crucial for the control of schistosomiasis and other snail-borne infections. The need to use plant s has received increased interest as an inexpensive technology because of the high cost of...
UMBILICAL CORD BLOOD BILIRUBIN AS A PREDICTOR FOR NEONATAL HYPERBILIRUBINEMIA
Jaundice is a clinical condition that is often present in pediatric practice and constitutes one of the major issues within the neonatal period. It occurs in both the physiological and pathological processes in newborns....
STUDY OF PHYTOCHEMICAL SCREENING, ANTIOXIDANT, ANTIDIARRHOEAL AND ANALGESIC ACTIVITIES OF HYDROMETHANOL EXTRACTS OF POLYALTHIA SUBEROSA (ROXB.)
Hydromethanol extracts of three different parts namely leaves, bark and fruits of the Polyalthia suberosa (Roxb.) was screen for it’s antioxidant, analgesic and antidiarrheal activity. Bark extracts showed the maximum r...
NEPHROTOXICITY: AN OVERVIEW
Nephrotoxicity is one of the most common situations faced by people due to easy availability of over the counter medication like NSAIDs, antibiotics and angiotensin converting enzyme inhibitors etc. There are various typ...
A STUDY FOR LINGUAL NERVE PROTECTION DURING EXTRACTION OF THIRD MOLAR USING LINGUAL REACTOR
Lingual nerve damage during surgical removal of third molar is most common nerve damage reported. Its incidence rate has been reported from 0.6 to as high as 22% in various studies conducted. Our aim was to study the eff...