Evaluation of non-culture diagnosis of invasive meningococcal disease by polymerase chain reaction (PCR)

• Published in: FEMS immunology and medical microbiology 2003-10, Vol.39 (1), p.31-36
• Main Authors: Tzanakaki, Georgina, Tsolia, Maria, Vlachou, Vasiliki, Theodoridou, Maria, Pangalis, Anastasia, Foustoukou, Maria, Karpathios, Themistocles, Blackwell, C.Caroline, Kremastinou, Jenny
• Format: Article
• Language: English
• Subjects: Bacteriological Techniques - statistics & numerical data; Base Sequence; Biological and medical sciences; Case-Control Studies; Child; Diagnosis; DNA, Bacterial - genetics; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Humans; insertion sequence IS1106; Meningococcal disease; Meningococcal Infections - diagnosis; Meningococcal Infections - microbiology; Microbiology; Neisseria meningitidis; Neisseria meningitidis - classification; Neisseria meningitidis - genetics; Neisseria meningitidis - isolation & purification; orf-2 gene; Polymerase chain reaction; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - statistics & numerical data; Sensitivity and Specificity; siaD gene
• ISSN: 0928-8244; 1574-695X
• DOI: 10.1016/S0928-8244(03)00175-5

Antibiotic treatment prior to transport or admission to hospital has reduced the proportion of cases of invasive meningococcal disease (IMD) from which Neisseria meningitidis can be isolated by standard microbiological techniques. Identification of meningococci by polymerase chain reaction (PCR) was assessed in relation to microbiological diagnosis for cases over a 4-year period between 1998 and 2001. A screening assay for the IS 1106 gene was used to detect meningococcal DNA and five additional assays for siaD and orf-2 genes were performed to determine the serogroup. PCR results were compared with results of bacteriological culture, other laboratory test results and clinical data. The sensitivity of the PCR assay for culture-confirmed cases was 98.5%. The specificity of the assay was 96% based on test results for patients from whom other bacteria were isolated, children with viral meningitis and afebrile negative controls. The siaD B/C/W-135 and Y as well as the orf-2 gene for serogroup A PCR assays were able to determine the serogroup for 75.2% of cases that were positive by PCR screening assay. When isolates from patients with IMD were tested by both agglutination and PCR, the results agreed in all cases. PCR is a useful tool for diagnosis of IMD when Gram stain and culture tests are negative due to antibiotic treatment prior to collection of samples for microbiological analyses.