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. 2015 Mar;92(3):583-7.
doi: 10.4269/ajtmh.14-0593. Epub 2014 Dec 15.

Recombinase polymerase amplification-based assay to diagnose Giardia in stool samples

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Recombinase polymerase amplification-based assay to diagnose Giardia in stool samples

Zachary Austin Crannell et al. Am J Trop Med Hyg. 2015 Mar.

Abstract

Giardia duodenalis is one of the most commonly identified parasites in stool samples. Although relatively easy to treat, giardiasis can be difficult to detect as it presents similar to other diarrheal diseases. Here, we present a recombinase polymerase amplification-based Giardia (RPAG) assay to detect the presence of Giardia in stool samples. The RPAG assay was characterized on the bench top using stool samples spiked with Giardia cysts where it showed a limit-of-detection nearly as low as the gold standard polymerase chain reaction assay. The RPAG assay was then tested in the highlands of Peru on 104 stool samples collected from the surrounding communities where it showed 73% sensitivity and 95% specificity against a polymerase chain reaction and microscopy composite gold standard. Further improvements in clinical sensitivity will be needed for the RPAG assay to have clinical relevance.

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Figures

Figure 1.
Figure 1.
The recombinase polymerase amplification-based Giardia (RPAG) assay consistently detects as few as 103–103.5 cysts per milliliter of stool during bench top testing. As described in the Methods section, the signal/background ratio (SBR) was calculated for each test line. The calculated SBR is shown below each strip. All samples with an SBR > 1.07 were considered positive.
Figure 2.
Figure 2.
The recombinase polymerase amplification-based Giardia (RPAG) assay only tests positive for samples containing DNA extracted from Giardia. As described in the Methods section, the signal/background ratio (SBR) was calculated for each test line. The calculated SBR is shown to the right of each strip. All samples with an SBR > 1.07 were considered positive.

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