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PLoS Negl Trop Dis. 2012;6(12):e1948. doi: 10.1371/journal.pntd.0001948. Epub 2012 Dec 13.

Diagnosis of brugian filariasis by loop-mediated isothermal amplification.

Author information

1
New England Biolabs, Ipswich, Massachusetts, United States of America.

Abstract

In this study we developed and evaluated a Brugia Hha I repeat loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Brugia genomic DNA. Amplification was detected using turbidity or fluorescence as readouts. Reactions generated a turbidity threshold value or a clear visual positive within 30 minutes using purified genomic DNA equivalent to one microfilaria. Similar results were obtained using DNA isolated from blood samples containing B. malayi microfilariae. Amplification was specific to B. malayi and B. timori, as no turbidity was observed using DNA from the related filarial parasites Wuchereria bancrofti, Onchocerca volvulus or Dirofilaria immitis, or from human or mosquito. Furthermore, the assay was most robust using a new strand-displacing DNA polymerase termed Bst 2.0 compared to wild-type Bst DNA polymerase, large fragment. The results indicate that the Brugia Hha I repeat LAMP assay is rapid, sensitive and Brugia-specific with the potential to be developed further as a field tool for diagnosis and mapping of brugian filariasis.

PMID:
23272258
PMCID:
PMC3521703
DOI:
10.1371/journal.pntd.0001948
[Indexed for MEDLINE]
Free PMC Article

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