Loop-Mediated Isothermal Amplification Assay for Detection of Haemophilus influenzae Type b in Cerebrospinal Fluid
- Authors
- Kim, Dong Wook; Kilgore, Paul Evan; Kim, Eun Jin; Kim, Soon Ae; Dang Duc Anh; Seki, Mitsuko
- Issue Date
- Oct-2011
- Publisher
- American Society for Microbiology
- Keywords
- REAL-TIME PCR; STREPTOCOCCUS-PNEUMONIAE; BACTERIAL-MENINGITIS; GENE; DNA; DISEASE BURDEN; VACCINE; LAMP; CHILDREN
- Citation
- Journal of Clinical Microbiology, v.49, no.10, pp 3621 - 3626
- Pages
- 6
- Indexed
- SCI
SCIE
SCOPUS
- Journal Title
- Journal of Clinical Microbiology
- Volume
- 49
- Number
- 10
- Start Page
- 3621
- End Page
- 3626
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/37171
- DOI
- 10.1128/JCM.00515-11
- ISSN
- 0095-1137
1098-660X
- Abstract
- Haemophilus influenzae type b (Hib) is one of the leading causes of meningitis in developing countries. To establish and evaluate a novel loop-mediated isothermal amplification (LAMP) assay for Hib, we designed a LAMP primer set targeting the Hib-specific capsulation locus. LAMP detected 10 copies of purified DNA in a 60-min reaction. This indicated that the detection limit of LAMP was > 100-fold lower than the detection limits of both a PCR for the detection of bexA and a nested PCR for Hib (Hib PCR). No H. influenzae, other than Hib or control bacteria, was detected. Linear determination ranged from 10 to 1,000,000 microorganisms per reaction mixture using real-time turbidimetry. We evaluated the Hib LAMP assay using a set of 52 randomly selected cerebrospinal fluid (CSF) specimens obtained from children with suspected meningitis. For comparison, the CSF specimens were tested using a conventional Hib PCR assay. Hib was detected in 30 samples using LAMP and in 22 samples using the Hib PCR assay. The Hib PCR showed a clinical sensitivity of 73.3% and a clinical specificity of 100% relative to the Hib LAMP assay. These results suggest that further development and evaluation of the Hib LAMP will enhance the global diagnostic capability for Hib detection.
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