Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea

Show full item record



Permalink

http://hdl.handle.net/10138/311365

Citation

Mero , S , Kirveskari , J , Antikainen , J , Ursing , J , Rombo , L , Kofoed , P-E & Kantele , A 2017 , ' Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea ' , Infectious Diseases , vol. 49 , no. 9 , pp. 655-663 . https://doi.org/10.1080/23744235.2017.1320728

Title: Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea
Author: Mero, Sointu; Kirveskari, Juha; Antikainen, Jenni; Ursing, Johan; Rombo, Lars; Kofoed, Poul-Erik; Kantele, Anu
Contributor organization: Department of Diagnostics and Therapeutics
Clinicum
Anu Kantele-Häkkinen Research Group
Department of Medicine
University of Helsinki
Infektiosairauksien yksikkö
HUS Inflammation Center
Date: 2017
Language: eng
Number of pages: 9
Belongs to series: Infectious Diseases
ISSN: 2374-4235
DOI: https://doi.org/10.1080/23744235.2017.1320728
URI: http://hdl.handle.net/10138/311365
Abstract: Background: In developing countries, diarrhoea is the most common cause of death for children under five years of age, with Giardia lamblia, Cryptosporidium and Entamoeba histolytica as the most frequent pathogenic parasites. Traditional microscopy for stool parasites has poor sensitivity and specificity, while new molecular methods may provide more accurate diagnostics. In poor regions with sample storage hampered by uncertain electricity supply, research would benefit from a method capable of analysing dried stools. Methods: A real-time multiplex PCR method with internal inhibition control was developed for detecting Giardia lamblia, Cryptosporidium hominis/parvum and Entamoeba histolytica directly from stool specimens. Applicability to dried samples was checked by comparing with fresh ones in a small test material. Finally, the assay was applied to dried specimens collected from Guinea-Bissauan children with diarrhoea. Results: The PCR's analytical sensitivity limit was 0.1 ng/ml for G. lamblia DNA, 0.01 ng/ml for E. histolytica DNA and 0.1 ng/ml for Cryptosporidium sp. In the test material, the assay performed similarly with fresh and dried stools. Of the 52 Guinea-Bissauan samples, local microscopy revealed a parasite in 15%, while PCR detected 62% positive for at least one parasite: 44% of the dried samples had Giardia, 23% Cryptosporidium and 0% E. histolytica. Conclusions: Our new multiplex real-time PCR for protozoa presents a sensitive method applicable to dried samples. As proof of concept, it worked well on stools collected from Guinea-Bissauan children with diarrhoea. It provides an epidemiological tool for analysing dried specimens from regions poor in resources.
Subject: Real-time PCR
Giardia
Cryptosporidium
Entamoeba
aetiology
developing country
children
travel
diarrhoea
parasite
stool parasite
INTESTINAL PARASITIC INFECTIONS
HUMAN GASTROINTESTINAL-TRACT
REAL-TIME PCR
DEVELOPING-COUNTRIES
FILTER-PAPER
DIAGNOSIS
PROTOZOA
HEALTH
ASSOCIATION
DUODENALIS
3121 General medicine, internal medicine and other clinical medicine
Peer reviewed: Yes
Usage restriction: openAccess
Self-archived version: acceptedVersion


Files in this item

Total number of downloads: Loading...

Files Size Format View
Mero_et_al._201 ... ted_11.4.2017_Inf.Dis..pdf 180.8Kb PDF View/Open

This item appears in the following Collection(s)

Show full item record