Molecular Detection of the Carriage Rate of Four Intestinal Protozoa with Real-Time Polymerase Chain Reaction: Possible Overdiagnosis of Entamoeba histolytica in Nigeria

• Published in: The American journal of tropical medicine and hygiene 2015-08, Vol.93 (2), p.257-262
• Main Authors: Efunshile, Michael A, Ngwu, Bethrand A F, Kurtzhals, Jørgen A L, Sahar, Sumrin, König, Brigitte, Stensvold, Christen R
• Format: Article
• Language: English
• Subjects: Adolescent; Carrier State - epidemiology; Carrier State - parasitology; Child; Cross-Sectional Studies; Cryptosporidium; Cryptosporidium - classification; Cryptosporidium - isolation & purification; Diarrhea - epidemiology; Diarrhea - parasitology; DNA, Protozoan - isolation & purification; Entamoeba - classification; Entamoeba - isolation & purification; Entamoeba histolytica; Entamoeba histolytica - classification; Entamoeba histolytica - isolation & purification; Entamoebiasis - diagnosis; Entamoebiasis - epidemiology; Feces - parasitology; Female; Giardia; Giardia - classification; Giardia - isolation & purification; Giardia intestinalis; Giardiasis - diagnosis; Giardiasis - epidemiology; Healthy Volunteers; Humans; Male; Nigeria - epidemiology; Prevalence; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity; Socioeconomic Factors; Surveys and Questionnaires
• ISSN: 0002-9637; 1476-1645
• DOI: 10.4269/ajtmh.14-0781

Diarrhea remains the second largest killer of children worldwide, and Nigeria ranks number two on the list of global deaths attributable to diarrhea. Meanwhile, prevalence studies on potentially diarrheagenic protozoa in asymptomatic carriers using molecular detection methods remain scarce in sub-Saharan countries. To overcome sensitivity issues related to microscopic detection and identification of cysts in stool concentrates, real-time polymerase chain reaction (PCR) was used to analyze genomic DNAs extracted from stool samples from 199 healthy school children for Entamoeba histolytica, E. dispar, Giardia intestinalis, and Cryptosporidium. Questionnaires were administered for epidemiological data collection. E. histolytica was not detected in any of the samples, whereas Giardia (37.2%), E. dispar (18.6%), and Cryptosporidium (1%) were found. Most of the children sourced their drinking water from community wells (91%), while the majority disposed of feces in the bush (81.9%). Our study is the first to use real-time PCR to evaluate the epidemiology of E. histolytica, Giardia, and Cryptosporidium in Nigeria where previous studies using traditional diagnostic techniques have suggested higher and lower carriage rates of E. histolytica and Giardia, respectively. It is also the first study to accurately identify the prevalence of common potentially diarrheagenic protozoa in asymptomatic carriers in sub-Saharan Africa.