Chika Takano, Japan
Nihon University MedicineAuthor Of 1 Presentation
MENINGOCOCCAL AND PNEUMOCOCCAL TYPING METHODS OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (ID 641)
Abstract
Background
We established meningococcal and pneumococcal typing methods of loop-mediated isothermal amplification (LAMP) and assessed their suitability for detecting meningococcal and pneumococcal types in cerebrospinal fluid (CSF).
Methods
The LAMP assays were developed to identify six major serogroups (A, B, C, X, Y, and W-135) of meningococcus and the 13 capsular types that are included in PCV13 (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F). We evaluated test reactivity, specificity, sensitivity, and compared the results between established LAMP and conventional PCR assays. To evaluate clinical performance of these assays, randomly selected CSF from children with suspected meningitis collected between 1998 and 2002 in Vietnam, China and Korea.
Results
The specificity of the primer sets was confirmed, and the assays could detect ten to 100 copies of purified DNA. The 31 meningococcal LAMP positive specimens were analyzed by serogrouping LAMP. The LAMP assay identified serogroups of 29 of the Neisseria meningitidis in CSF. The detection limits of the pneumococcal serotype specific LAMP assay were comparable when using DNA-spiked CSF, as compared to using purified DNA as the template.
Conclusions
Highly accurate LAMP assays have been developed for meningococcal and pneumococcal typing for globally use of surveillance and vaccine effectiveness studies.