A METHOD FOR DETERMINATION OF NEISSERIA MENINGITIDIS SEROGROUP A, B, C AND W BY REAL-TIME PCR


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Abstract

Aim. Deve1opment and testing of a rea1-time PCR method for detection of Neisseria meningitidis serogroup A, B, C and W DNA. Materials and methods. Reference strains and 187 samp1es of cerebrospina1 f1uid (CSF) from meningococci meningitis patients were used in the study. Mu1tip1ex PCR was carried out in an instrument with 5 channe1s of fluorescent detection. Results. Ana1ysis of specific serogroup 1oci of the genome and design of o1igonuc1eotides for the detection of DNA of a11 the capsu1e meningococci and 4 serogroups in particu1ar was carried out. PCR conditions were optimized; specificity was shown and ana1ytica1 sensitivity was eva1uated using reference strains. DNA of the fo11owing serogroups was detected during study of c1inica1 CSF samp1es: A - in 103 samp1es (55%), B - in 45 (24%), C - in 30 (16%), W - in 5 (3%). On1y DNA of meningococci capsu1e gene ctrA was found in 4 samp1es; presumab1y, they contained DNA of other serogroups. Mu1ti1ocus sequence-typing and detection of antigenic determinants of PorA and FetA genes for 27 DNA samp1es of group A menincococci as we11 as DNA of 5 group W meningococci and 4 ungroupab1e was carried out. Conclusion. The method proposed a11ows to carry out serogrouping of no less than 95% of strains or DNA samples isolated from CSF of meningococci infection patients. Combined with other recommended non-cultural methods of genotyping, it may be useful for complex characteristics of pathogenic meningococci.

About the authors

K. O Mironov

Centra1 Research Institute of Epidemio1ogy, Moscow, Russia

A. E Platonov

Centra1 Research Institute of Epidemio1ogy, Moscow, Russia

O. P Dribnokhodova

Centra1 Research Institute of Epidemio1ogy, Moscow, Russia

V. I Kuseva

Centra1 Research Institute of Epidemio1ogy, Moscow, Russia

G. A Shipulin

Centra1 Research Institute of Epidemio1ogy, Moscow, Russia

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Copyright (c) 2014 Mironov K.O., Platonov A.E., Dribnokhodova O.P., Kuseva V.I., Shipulin G.A.

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