Background and Aim : Cystic fibrosis (CF), is the most common fatal multisystem genetic disease in Caucasian populations, results from mutations in the CF transmembrane conductance regulator (CFTR) gene and affects almost all of the body’s function exocrine glands. This causes the secretion of thick and sticky mucus in the lungs, which leads to narrowing of the airways and, as a result, traps of viruses, bacteria, and fungal spores in the air in this mucus, leading to various infections. CF Patients have chronic and recurrent lung infections, which is why lung problems are often the leading cause of death in those with the disease. In this study, a multiplex PCR method was developed to identify Haemophilus influenzae non-typeable, Burkholderia cepacia complex, and Pseudomonas aeruginosa, in oropharyngeal samples from CF patients.Methods : Forty-four oropharyngeal samples were collected from CF patients (۲۵ male, and ۱۹ female) in Children's Medical Center Hospital. Samples were studied by culture and multiplex PCR method. SDS and chloroform-isoamyl alcohol were used for cell lysis and DNA purification.
Multiplex PCR was done using three pairs of primers targeting specific genomic sequences of each species.Results :
Multiplex PCR results revealed ۷۲.۲ % P. aeruginosa and ۳۶.۴ % B. cepacia complex, as well as ۲۲.۷ % of H. influenzae.
Multiplex PCR was able to detect CF patients cloned with P. aeruginosa who were not identified by standard culture.
Multiplex PCR detected P. aeruginosa in ۲۱ participants, H. influenzae in ۱۰ participants, and B. cepacia complex (BCC) in ۱۶ participants, that were not detected in culture-dependent technique. Comparing to culture results, the sensitivity and specificity values of multiplex PCR for P. aeruginosa identification were, respectively, ۱۰۰% and ۳۶.۴%. Specificity values of multiplex PCR for BCC and H. influenzae were ۱۰۰%. The sensitivity of the multiplex PCR for B. cepacia complex and H. influenzae cannot be assessed because no positive results have been recorded for these two bacteria by culture method. Conclusion : we conclude that multiplex PCR provides a rapid method for detecting these ۳ bacterial species in CF patients’ samples with greater accuracy and sensitivity than the culture method