Design of primers for pertussis diagnosis by Real Time PCR and determination of its sensitivity and specificity in comparison with commercial kits.

Abstract

Background and Aim: Pertussis vaccination in this country has been going on for many years and active infection or vaccination will provide immunity in 85% of cases. However, every 2-5 years outbreaks in unprotected adults creates an epidemy for children and infants. Based on conserved genomic sequences, Real time PCR could be an easy, cost- benefit, fast and highly sensitive method for pertussis detection. Materials and Methods: A total of 170 nasopharyngeal swabs of infants with history of cough for more than 2 weeks were collected. In the first stage, Bordetella pertussis bacteria detection was performed by culture and followed by Real time PCR using a commercial kit and then repeated with newly designed primers. Results: Performance of our home made primers for detecting pertussis using Real Time PCR in comparison with those by commercial kit was acceptable based on diagnostic classical guidance (WHO) and the (CDC). Conclusions: Real time PCR test with new primers in comparison with culture techniques is more suitable, high sensitivity and can provide more informative values for pertussis detection.