Loop Mediated Isothermal Amplification (LAMP) assay for Rapid detection of Streptococcus agalactiae (Group B Streptococcus - GBS) in vaginal swabs - A Proof of Concept Study

J. McKenna, C. Cox, D. J. Fairley, R. Burke, M. D. Shields, A. Watt, P. V. Coyle

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)
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Abstract

Purpose: Neonatal sepsis caused by Streptococcus agalactiae [group B streptococcus (GBS)] is a life-threatening condition, which is preventable if colonized mothers are identified and given antibiotic prophylaxis during labour. Conventional culture is time consuming and unreliable, and many available non-culture diagnostics are too complex to implement routinely at point of care. Loop-mediated isothermal amplification (LAMP) is a method that, enables the rapid and specific detection of target nucleic acid sequences in clinical materials without the requirement for extensive sample preparation.

Methodology: A prototype LAMP assay targeting GBS sip gene is described.

Results: The assay was 100 % specific for GBS, with a limit of detection of 14 genome copies per reaction. The clinical utility of the LAMP assay for rapid direct molecular detection of GBS was determined by testing a total of 157 vaginal swabs with minimal sample processing using a rapid lysis solution. Compared to a reference quantitative real-time PCR assay, the direct LAMP protocol had a sensitivity and specificity of 95.4 and 100 %, respectively, with positive and negative predictive values of 100 and 98.3 %, respectively. Positive and negative likelihood ratios were infinity and 0.05, respectively. The direct LAMP method required a mean time of 45 min from the receipt of a swab to generation of a confirmed result, compared to 2 h 30 min for the reference quantitative real-time PCR test.

Conclusion: The direct LAMP protocol described is easy to perform, facilitating rapid and accurate detection of GBS in vaginal swabs. This test has a potential for use at point of care.
Original languageEnglish
Pages (from-to)294-300
JournalJournal of Medical Microbiology
Volume66
Issue number3
Early online date23 Mar 2017
DOIs
Publication statusPublished - Mar 2017

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