Real-time polymerase chain reaction detection of Neisseria meningitidis in formalin-fixed tissues from sudden deaths.
Accurate identification of meningococcal sudden deaths is needed to avoid underestimation of the true incidence of the disease. This study analyzed the usefulness of a real-time polymerase chain reaction (PCR) protocol using MGB (3'-minor groove binder) probes to detect Neisseria meningitidis in formalin-fixed paraffin-embedded tissues from sudden deaths where a meningococcal fulminating infection was suspected. The protocol included detection of meningococcal DNA (ctrA gene), multiplex B/C PCR serogrouping (siaD gene), and rapid confirmation of PCR products by microcapillary electrophoresis. Sixty-nine tissues from 15 culture-confirmed meningococcal sudden deaths were analyzed (positive cases). Validation studies were performed. In each positive case, both the ctrA and the B/C siaD genes were detected. The ctrA was detected in 81.2% of the samples, whereas the serogroup (B or C) was identified in 44.9% of them. Therefore, this protocol may improve nonculture diagnosis and case ascertainment in meningococcal disease deaths, particularly when formalin-fixed tissues are the only available specimen.
Fernández-Rodríguez
Amparo
A
Microbiology Laboratory, Instituto Nacional de Toxicología y Ciencias Forenses, c) Luis Cabrera no 9, 28002 Madrid, Spain. a.fernandez@mju.es
Alcalá
Belén
B
Alvarez-Lafuente
Roberto
R
D000293:Adolescent; D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D003645:Death, Sudden / Q000209:etiology*; D019075:Electrophoresis, Capillary; D005260:Female; D005838:Genotype; D006801:Humans; D007223:Infant; D008297:Male; D008589:Meningococcal Infections / Q000175:diagnosis*; D008875:Middle Aged; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D017931:DNA Primers