Development of a rapid, simple multiplex PCR-dipstick assay for the detection of Neisseria meningitidis serogroups in clinical isolates

The rapid identification and serogrouping of <i>Neisseria meningitidis</i> are crucial for effectively managing invasive meningococcal disease (IMD) patients and their close contacts, particularly in developing countries with limited laboratory resources. We developed a simple multiplex polymerase chain reaction (PCR)-dipstick DNA chromatography (mPCR-dipstick) assay for detecting six major serogroups (A, B, C, W, Y, and X) prevalent worldwide. The assay performance, sensitivity, and specificity were evaluated using 116 unencapsulated and encapsulated <i>N. meningitidis</i>, 29 other <i>Neisseria</i> spp., and 11 non-<i>Neisseria</i> spp. strains. The mPCR-dipstick assay successfully identified all unencapsulated and encapsulated <i>N. meningitidis</i> strains and accurately differentiated the six major serogroups. The detection limit was 4.1 × 10⁴ genome copies and 5.3-266 colony-forming units (CFU) per reaction, indicating 100% sensitivity and 100% specificity. This convenient, sensitive, and rapid assay provides considerable advantages for diagnosing, treating, and controlling meningococcal infections in resource-limited settings, especially in high IMD endemic regions, including West Africa.