Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identification of Giardia intestinalis , Cryptosporidium parvum / Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review

Laude, Adrien; Valot, Stéphane; Désoubeaux, Guillaume; Argy, Nicolas; Nourrisson, Céline; Pomarès, Christelle; Machouart, M.; Govic, Yohann Le; Dalle, Frédéric; Botterel, Françoise; Bourgeois, Nathalie; Cateau, Estelle; Leterrier, Marion; Pape, Patrice Le; Morio, Florent

doi:10.1016/j.cmi.2015.10.019

Cited by 65 publications

(71 citation statements)

References 30 publications

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“…Therefore, alternative techniques have been evaluated to overcome limitations of the conventional microscopy-based diagnostic methods in order to achieve more accurate and specific diagnosis. In recent years there has been greater attention given to DNA-based diagnostic methods, including conventional and real-time PCR-based diagnostic modalities (Laude et al, 2016). However, stool-based PCR assays require specialized equipment, well-equipped diagnostic laboratories, specialized training and skilled staff.…”

Section: Discussionmentioning

confidence: 99%

Stool antigen immunodetection for diagnosis of Giardia duodenalis infection in human subjects with HIV and cancer

Nooshadokht

Kalantari-Khandani

Sharifi

et al. 2017

Journal of Microbiological Methods

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Human infection with the protozoan parasite Giardia duodenalis is one the most common parasitic diseases worldwide. Higher incidence rates of giardiasis have been reported from human subjects with multiple debilitating chronic conditions, including hypogammaglobulinemia and common variable immunodeficiency (CVID). In the current study, stool specimens were collected from 199 individuals diagnosed with HIV or cancer and immunocompetent subjects. The sensitivity of microscopy-based detection on fresh stool preparations, trichrome staining and stool antigen immunodetection for the diagnosis of G. duodenalis were 36%, 45.5% and 100%, respectively when compared with a highly sensitive stool-based PCR method as the gold standard. Further multilocus molecular analyses using glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) demonstrated that the AI genotype of G. duodenalis was the most prevalent, followed by the AII genotype and mixed (AI+B) infections. We concluded that stool antigen immunodetection-based immunoassays and stool-based PCR amplification had comparable sensitivity and specificity for the diagnosis of G. duodenalis infections in these populations. Stool antigen detection-based diagnostic modalities are rapid and accurate and may offer alternatives to conventional microscopy and PCR-based diagnostic methods for the diagnosis of G. duodenalis in human subjects living with HIV or cancer.

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Section: Discussionmentioning

confidence: 99%

Stool antigen immunodetection for diagnosis of Giardia duodenalis infection in human subjects with HIV and cancer

Nooshadokht

Kalantari-Khandani

Sharifi

et al. 2017

Journal of Microbiological Methods

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“…Those diagnostic accuracy tests indicated a relatively good value as a direct method for diagnosis of cryptosporidiosis among diarrheic children. Corroborating to that reported by Laude et al,(2016) (36) microscopy is the reference standard for routine laboratory diagnosis in faecal parasitology, but there is a growing need for alternative methods to overcome the limitations of microscopic examination, e.g. the high dependence on expertise of the operator.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Different Diagnostic Approaches for Detection of Cryptosporidium in Stools of Diarrheic Children

Eassa

Flefel²,

El-Masry

et al. 2017

Journal of High Institute of Public Health

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Background: Cryptosporidiosis is of utmost importance especially in vulnerable age groups in developing countries. Malnourished children are more susceptible to recurrent diarrheal episodes, which can lead to chronic nutritional and cognitive sequelae or even death. Objective(s): to evaluate four different diagnostic approaches for Cryptosporidium infection in stools of diarrheic children. Methods:One hundred stool specimens were collected from diarrheic children in Alexandria University Children Hospital (El-Shatby). All samples were investigated by four techniques; directly by microscopic detection of Cryptosporidium oocysts using modified Ziehl-Neelsen (MZN) stain. Indirectly, through detection of coproantigen by enzyme linked immunosorbent assay (ELISA) and rapid strip test. Cryptosporidium DNA was detected by conventional polymerase chain reaction (PCR). Results: Using the four methods, 65% of examined children had Cryptosporidium infection, while Cryptosporidium oocysts were shown by MZN stain technique among 41%. However, by rapid strip test, ELISA, and PCR the percentages were 45%, 48%, and 59%, respectively. PCR elicited the highest diagnostic efficiency (64%) among the three diagnostic non-microscopic techniques when the MZN technique was used as the gold standard test. However, rapid strip test showed the least diagnostic efficiency (48%) when compared to PCR that was considered as the gold standard test. Meanwhile, ELISA was of moderate performance when compared to either PCR or to MZN technique used as gold standard test. Conclusion: PCR was more sensitive than rapid strip test and ELISA. It is time saving, but not cost effective. The rapid strip test could be considered as a complementary (additional) tool rather than a substitute for microscopic examination. It could be used for screening in cases of outbreaks of diarrhea for faster management of the problem.

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“…Nowadays, the highest sensitivity and specificity (nearly 100%) for E. histolytica infection diagnosis is achieved with real-time PCR [17]. Based on its high specificity, we were able to delimit the 25 samples (9.2%) positive for Entamoeba spp.…”

Section: Discussionmentioning

confidence: 84%

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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Cited by 65 publications

References 30 publications

Stool antigen immunodetection for diagnosis of Giardia duodenalis infection in human subjects with HIV and cancer

Stool antigen immunodetection for diagnosis of Giardia duodenalis infection in human subjects with HIV and cancer

Evaluation of Different Diagnostic Approaches for Detection of Cryptosporidium in Stools of Diarrheic Children

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

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