A Pentaplex Real-Time Polymerase Chain Reaction Assay for Detection of Four Species of Soil-Transmitted Helminths

Abstract

Soil-transmitted helminth infections remain a major public health burden in low- and middle-income countries. The traditional diagnosis by microscopic examination of fecal samples is insensitive and time-consuming. In this study, a pentaplex real-time polymerase chain reaction (PCR) was evaluated for the simultaneous detection of Ancylostoma, Necator americanus, Ascaris lumbricoides, and Strongyloides stercoralis. The results were compared with those obtained by conventional parasitological diagnostic methods. Real-time PCR was positive in 48 of 77 samples (62.3%) and microscopic examination was positive in six samples (7.8%) only (P < 0.05). In conclusion, the real-time PCR assay described in this study provides a specific and sensitive diagnostic tool for the detection of these four helminth species in epidemiological studies and monitoring of treatment programs.

A Novel, Multi-Parallel, Real-Time Polymerase Chain Reaction Approach for Eight Gastrointestinal Parasites Provides Improved Diagnostic Capabilities to Resource-Limited At-Risk Populations

“…8 The qPCR has been useful in the diagnosis of pathogenic microbes causing tissue infections, including amebic liver abscess, and has also been shown to have a high degree of sensitivity and specificity in the detection of enteric pathogens. 2,3,9 As with other real-time PCR systems, to broaden the spectrum of enteric pathogen detection, a simple to execute, multi-parallel qPCR approach was developed that enables detection of an unlimited number of a parasite species provided that sequence information is available.…”

“…With the development of newer multiplex polymerase chain reaction (PCR)-based assays in stool for pathogenic parasite (hookworm, Strongyloides stercoralis, Ascaris lumbricoides, Entamoeba histolytica, Cryptosporidium parvum, and Giardia lamblia) identification, it is likely that gastrointestinal parasite identification in the stool will become more sensitive and objective. [1][2][3] Classically, the diagnosis of intestinal parasites depends on the life cycle of the parasite and its egg/larvae shedding patterns. Although microscopy and quantitative real-time PCR (qPCR) are dependent on the presence of parasite material in the sample, 4 qPCR is less subjective and for several parasites (e.g., E. histolytica) more specific.…”

Nested pcr based diagnosis of trichuriasis and appraisal of primer sensitivity and specificity for reliable detection in clinical samples

“…A prevalence rate of 70.0% T. trichiura and 10.7% T. vulpis were observed in a rural community in northwestern Thailand using molecular technique (Areekul et al 2010). Studies conducted across the world revealed the significance of molecular diagnosis of SoilTransmitted Helminths over microscopy methods (Basuni et al 2011;Arndt et al 2013).…”

Application of a real-time PCR method for detecting and monitoring hookworm Necator americanus infections in Southern China

“…However, methods based on the internal transcribed spacers (ITS-1 and 2) of the nuclear ribosomal DNA (rDNA) have received the widest applications because of their lower mutation rates within species and substantial differences between species [13,14] . Species-specific PCR primers have been designed to differentiate N. americanus from A. duodenale, and the method proved useful in epidemiological survey of hookworm infections [15,16] . In recent years, quantitative real-time PCR has been used in parasitology to monitor the prevalence of geohelminths [17,18] .…”