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Optimization of LipL32 PCR assay for increased sensitivity in diagnosing leptospirosis.

Abstract：

:Early diagnosis of leptospirosis in humans is critical with regard to initiation of appropriate treatment; however, the gold standard serological test cannot detect antibodies until nearly a week after symptom onset. PCR has been shown to be sensitive and specific in the early phase of leptospirosis. Previously, we developed and validated a TaqMan PCR assay targeting lipL32. We reoptimized and validated this assay using PerfeCTa® qPCR ToughMix®, Low ROX™ (Quanta Biosciences, Gaithersburg, MD, USA). For optimization with the new mix, the final primer concentrations were increased from 0.5 μmol/L to 0.9 μmol/L compared to our previous assay, and the probe concentration increased from 0.1 μmol/L to 0.125 μmol/L. This newly optimized assay resulted in a lower limit of detection and increased diagnostic sensitivity. Here, we present the performance data of the improved assay and describe several clinical cases that were initially negative but tested positive using the optimized assay.

journal_name

Diagn Microbiol Infect Dis

journal_title

Diagnostic microbiology and infectious disease

authors

Galloway RL,Hoffmaster AR

doi

10.1016/j.diagmicrobio.2015.03.024

subject

Has Abstract

pub_date

2015-07-01 00:00:00

pages

199-200

issue

3

eissn

0732-8893

issn

1879-0070

pii

S0732-8893(15)00116-9

journal_volume

82

pub_type

杂志文章

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