Document Type : Original Article
MOLECULAR BIOLOGY RESEARCH CENTER, BAQIYATALLAH (A.S) UNIVERSITY OF MEDICAL SCIENCES, TEHRAN, IRAN
Introduction: The aim of this study was to setup, optimize and introduce a sensitive and specific PCR detection method for identification of Neisseria meningitidis DNA in clinical samples.
Material and Methods: Capsular transport gene A (ctrA) was selected as a specific target sequence. This primer pair amplifies 101bp of the target gene. Neisseria meningitidis strain: ATCC; 13090 and Neisseria meningitides serogroup C were used as a standard organism for optimization experiments. A range of bacterial pathogens were used for specificity testing, including, Haemophilus influenzae Type b: ATCC; 49766, Escherichia coli: ATCC;35218, Enterobacter, Klebsiella pneumoniae, Streptococcus pneumoniae, Staphylococcus aureus and Streptococcus group D. Phenol – chloroform method was used for DNA extraction. Amplified product was detected by gel agarose electrophoresis, stained by ethidiome bromide.
Results: Our results confirmed amplification of the expected product. Specificity test proved no cross reaction with tested organisms. Sensitivity test detected 500fg of Neisseria meningitidis DNA as a final detection limit.
Conclusion: As a conclusion, PCR is a method with high sensivity and specificity and specificity which can be performed within 3 hours, and therefore utilization of this test in the clinical laboratories can help rapid diagnosis of Neisseria meningitides in clinical samples.