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

Mejia, Rojelio; Vicuña, Yosselin; Broncano, Nely; Sandoval, Carlos; Vaca, Maritza; Chico, Martha; Cooper, Philip J.; Nutman, Thomas B.

doi:10.4269/ajtmh.12-0726

Cited by 239 publications

(342 citation statements)

References 26 publications

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“…In developing countries and resource-limited areas, microscopy is the routine test for parasite infection diagnosis due to its low cost [16]. Compared to immunoassay, microscopy has a specificity of 97% and sensitivity of only 20% for E. histolytica/E.…”

Section: Discussionmentioning

confidence: 99%

Use of real-time polymerase chain reaction to identify Entamoeba histolytica in schoolchildren from northwest Mexico

Aguayo-Patrón

Castillo-Fimbres

Quihui‐Cota

et al. 2017

J Infect Dev Ctries

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Introduction: Entamoeba histolytica, E. dispar, and E. moshkovskii are morphologically identical, but intestinal amebiasis is caused only by E. histolytica. Mexico is among the countries with high amebae infection rates, although the contribution of pathogenic amoeba to the total detected cases remains unknown, especially in the northwestern dry region. Therefore, the aim of this study was to identify the actual prevalence of E. histolytica using real-time polymerase chain reaction (PCR) in schoolchildren of northwestern Mexico. Methodology: Participants were children from five public elementary schools in the low-socioeconomic-level suburban areas of Hermosillo, Sonora, Mexico. One stool sample was collected from each child and analyzed by the Faust technique for Entamoeba spp. and by real-time PCR for E. histolytica. Results: Analysis of stool samples from 273 children (9.0 ± 1.5 years of age) resulted in 25 (9.2%) positive for E. histolytica/E. dispar/E. moshkovskii by the Faust technique; of these, 3 were positive for E. histolytica by real-time PCR. In addition, 2 samples that were negative for E. histolytica/E. dispar/E. moshkovskii by the Faust technique were positive by real-time PCR. Conclusions: The actual prevalence of E. histolytica in our study population was 1.8%, which is lower than those reported in previous studies in other Mexican regions.

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Section: Discussionmentioning

confidence: 99%

Use of real-time polymerase chain reaction to identify Entamoeba histolytica in schoolchildren from northwest Mexico

Aguayo-Patrón

Castillo-Fimbres

Quihui‐Cota

et al. 2017

J Infect Dev Ctries

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“…The majority of Strongyloides PCR publications have used a real-time method with primers and probe published by Verweij et al 19 . This has also allowed for the development of multiplexed PCR 10,30,34,38 . Some studies evaluating the diagnostic accuracy of these PCR methods have used both morphological diagnosis and detection of PCR products as their reference standards, and are not reviewed here.…”

Section: Pcrmentioning

confidence: 99%

“…the 18S rRNA small subunit (SSU); the internal transcribed spacer region 1 (ITS-1); the 28S rRNA gene; or the cyclooxygenase gene (cox1) 19,[23][24][25][26][27][28][29][30][32][33][34][35][36][37] . Published sensitivities and specificities for…”

Section: Pcrmentioning

confidence: 99%

The laboratory diagnosis of Strongyloides stercoralis

Watts¹,

Robertson²,

Bradbury

2016

Microbiol. Aust.

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It is estimated that over 30million people worldwide are infected by the nematode, Strongyloides stercoralis1. It is endemic in sub-tropical and tropical parts of Australia, with high rates of infection documented in some indigenous communities2. Due to the potential for chronic autoinfection, that may persist for decades, migration leads to the presence of the infection in non-endemic areas1. Transmission to humans is generally through the penetration of larvae through the skin, following contact with faecally contaminated soil1. Disease severity ranges from asymptomatic chronic carriage to an overwhelming illness, where large numbers spread throughout the body, usually triggered by immunosuppression1.

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“…The amplicons were detected using the sequence FAM-CCC GCG GCG CTC GTC GCT AG-BHQ as probe. The DNA amplification programme was set up as previously describes by Mejia et al [24], as follows: after preheating at 60°C for 1 min, a cycle was performed at 95°C for 10 min followed by 45 cycles at 95°C during 15 s, 60°C for 1 min and a final extension at 60°C for 1 min.…”

Section: Real-time Pcr Analysismentioning

confidence: 99%

Evaluation of a Real-Time Polymerase Chain Reaction for the Laboratory Diagnosis of Giardia intestinalis in Stool Samples from Schoolchildren from the Centre-Ouest and Plateau Central Regions of Burkina Faso

Diagbouga¹,

Kientega²,

Erismann³

et al. 2017

Appli Micro Open Access

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Objective: Giardiasis, a zoonotic, diarrhoeal disease with worldwide occurrence, is routinely diagnosed by microscopic examination of stool samples. However, implementation of this method relies on skilled personal, it is time consuming and relatively low in sensitivity. A superior diagnostic approach to detect the causative agent Giardia intestinalis would, hence, be highly desirable. The current study aimed to assess real-time polymerase chain reaction (PCR) for the detection of G. intestinalis as an alternative to microscopy. Material and methods:Stool samples from healthy schoolchildren, aged 8-14 years, from eight schools of the Centre-Ouest and Plateau Central regions in Burkina Faso were collected within a cross-sectional study in February 2015. Microscopic examination was performed on two faecal samples collected over two consecutive days from 441 schoolchildren. Each faecal specimen was examined using Kato-Katz and formol-ether methods of concentration in addition to direct examination. Real-time PCR was used to detect G. intestinalis in all microscopy-positive and a random sample of microscopy-negative samples.Results: Microscopic examination revealed 94 microscopy-positive samples, and an overall G. intestinalis prevalence of 27.2%. Using the microscopic examination as the 'gold' standard, the overall sensitivity of real-time PCR was demonstrated to be 76.6% ranging from 58.3% to 94.1% and the specificity was 96.2% ranging from 96.2% to 100% across schools. Conclusion:Real-time PCR appears to be a solid detection method for G. intestinalis in the current setting. However, it needs to be further optimized to become a more sensitive tool for G. intestinalis diagnosis in low-income settings.

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

Cited by 239 publications

References 26 publications

Use of real-time polymerase chain reaction to identify Entamoeba histolytica in schoolchildren from northwest Mexico

Use of real-time polymerase chain reaction to identify Entamoeba histolytica in schoolchildren from northwest Mexico

The laboratory diagnosis of Strongyloides stercoralis

Evaluation of a Real-Time Polymerase Chain Reaction for the Laboratory Diagnosis of Giardia intestinalis in Stool Samples from Schoolchildren from the Centre-Ouest and Plateau Central Regions of Burkina Faso

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