This site needs JavaScript to work properly. Please enable it to take advantage of the complete set of features!
Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log in
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Jun;88(6):1041-7.
doi: 10.4269/ajtmh.12-0726. Epub 2013 Mar 18.

A novel, multi-parallel, real-time polymerase chain reaction approach for eight gastrointestinal parasites provides improved diagnostic capabilities to resource-limited at-risk populations

Affiliations

A novel, multi-parallel, real-time polymerase chain reaction approach for eight gastrointestinal parasites provides improved diagnostic capabilities to resource-limited at-risk populations

Rojelio Mejia et al. Am J Trop Med Hyg. 2013 Jun.

Abstract

Diagnosis of gastrointestinal parasites has traditionally relied on stool microscopy, which has low diagnostic sensitivity and specificity. We have developed a novel, rapid, high-throughput quantitative multi-parallel real-time polymerase chain reaction (qPCR) platform. Species-specific primers/probes were used for eight common gastrointestinal parasite pathogens: Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Giardia lamblia, Cryptosporidium spp., Entamoeba histolytica, Trichuris trichiura, and Strongyloides stercoralis. Stool samples from 400 13-month-old children in rural Ecuador were analyzed and the qPCR was compared with a standard direct wet mount slide for stool microscopy, as were 125 8-14-year-old children before and after anthelmintic treatment. The qPCR showed higher detection rates for all parasites compared with direct microscopy, Ascaris (7.0% versus 5.5%) and for Giardia (31.5% versus 5.8%). Using an enhanced DNA extraction method, we were able to detect T. trichiura DNA. These assays will be useful to refine treatment options for affected populations, ultimately leading to better health outcomes.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Correlation of parasite DNA concentration (fg/μL) with egg count by Kato-Katz method for Ascaris lumbricoides (top) (r = 0.713, P < 0.0001) and Trichuris trichiura (bottom) (r = 0.743, P < 0.0001) for 400 13-month-old-children, Ecuador. Only samples positive by high-throughput quantitative multi-parallel real-time polymerase chain reaction are shown.
Figure 2.
Figure 2.
Detection of polyparasitism by quantitative multi-parallel real-time polymerase chain reaction (qPCR) and microscopy in 400 13-month-old children, Ecuador. A, Proportion of stools positive for 0–4 parasites by either microscopy or qPCR. Asterisk indicates statistical significance (P < 0.05) difference in detection between microscopy and qPCR. B, Improved detection of the three most prevalent infections showing the distribution (in a Venn diagram) of these parasites based on microscopy or qPCR. Asterisk indicates a statistically significant (P < 0.05) difference in detection between microscopy and PCR.
Figure 3.
Figure 3.
Prevalence of Ascaris lunbricoides and Giardia lamblia before (pre) and 21 days after (post) albendazole/ivermectin treatment in 125 children 8–14 years of age, Ecuador. A. lumbricoides DNA decreased to undetectable levels post-treatment with the exception of two samples. A total of 35.1% of children remained positive for G. lamblia after empiric treatment.

References

    1. Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque AS, Ahmad N, Kirkpatrick BD, Houpt E, Snider C, Petri WA., Jr Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh. Clin Infect Dis. 2009;48:1191–1197. - PMC - PubMed
    1. ten Hove RJ, van Esbroeck M, Vervoort T, van den Ende J, van Lieshout L, Verweij JJ. Molecular diagnostics of intestinal parasites in returning travellers. Eur J Clin Microbiol Infect Dis. 2009;28:1045–1053. - PMC - PubMed
    1. Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, Rahumatullah A, Aziz FA, Zainudin NS, Noordin R. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg. 2011;84:338–343. - PMC - PubMed
    1. Pecson BM, Barrios JA, Johnson DR, Nelson KL. A real-time PCR method for quantifying viable Ascaris eggs using the first internally transcribed spacer region of ribosomal DNA. Appl Environ Microbiol. 2006;72:7864–7872. - PMC - PubMed
    1. Wolk DM, Schneider SK, Wengenack NL, Sloan LM, Rosenblatt JE. Real-time PCR method for detection of Encephalitozoon intestinalis from stool specimens. J Clin Microbiol. 2002;40:3922–3928. - PMC - PubMed

MeSH terms

LinkOut - more resources

Full text links
Sheridan PubFactory full text link Sheridan PubFactory Free PMC article
Cite

AltStyle によって変換されたページ (->オリジナル) /