Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt

Ghallab, Marwa M. I.; Aziz, Inas Z. Abdel; Shoeib, Eman Y.; El-Badry, Ayman A.

doi:10.1007/s12639-014-0601-9

Cited by 15 publications

(8 citation statements)

References 24 publications

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“…In the last decades in Egypt, there has been a reported varying Cryptosporidium preva-lence, a high of molecular prevalence rate of infection was shown in many studies up to 25% of the examined patients (El-Settawy and Fathy, 2012;Fathy et al, 2014;Ghallab et al, 2014). In contrast, much lower results (4.6%) reported (Abd El-Kader et al, 2011).…”

Section: Discussionmentioning

confidence: 96%

High-Resolution Melting Curve (Hrm) Analysis in Genotypic Discrimination of Cryptosporidium Isolates From Stool of Egyptian Children

Ghallab

Khater

Bouzid

et al. 2018

Journal of the Egyptian Society of Parasitology

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Cryptosporidiosis is a recognized child infectious killer and the second cause of diarrheal disease and death in infants. Assessing Cryptosporidium spp. genetic diversity is a real goal to elucidate its transmission dynamics and to design preventive measures in absence of effective treatment. Cryptosporidium isolates in stool of Egyptian children were detected using Acid Fast (AF) staining, copro-nPCR/RFLP assay and real time PCR high-resolution melting (HRM) curve analysis assay. Stool samples were collected from 335 children complaining of diarrhea and other GIT symptoms, attending the outpatient clinic of Abu El Reesh hospital, Kasr Al-Ainy School of Medicine, Cairo University. Two genotypes C. hominis and C. parvum were identified in 43 isolates from Egyptian children by copro-nPCR targeting COWP gene and HRM assay. Real time PCR HRM curve analysis, a closed-tube genotyping method, targeting ITS-2 gene confirmed the results of copro-nPCR/RFLP. It is simple, rapid, has more sample throughput, analysis capacities and data storage with less carry-over contamination and cost.

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Section: Discussionmentioning

confidence: 96%

High-Resolution Melting Curve (Hrm) Analysis in Genotypic Discrimination of Cryptosporidium Isolates From Stool of Egyptian Children

Ghallab

Khater

Bouzid

et al. 2018

Journal of the Egyptian Society of Parasitology

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“…thus increasing the potential for misdiagnosis [24][25][26] . Cryptosporidium ICT test showed good specificity (95%), however, it had a low sensitivity (54.5%).…”

Section: Discussionmentioning

confidence: 99%

Cryptosporidium spp. and Helicobacter pylori in a hospitalbased study of diarrheic immunocompromised Egyptian children: Insight into risk factors, and co-infection

Ibrahim

Ali

Abdel-Aziz

et al. 2022

Parasitologists United Journal

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Background: Immunocompromised children are more susceptible to a diversity of pathogens including norovirus, rotavirus, diarrheagenic Escherichia coli, Cryptosporidium spp. and Helicobacter pylori.Objective: This study aimed to determine the detection rate of H. pylori and Cryptosporidium spp.; their co-infection in a hospital-based study of diarrheic immunocompromised children and their predictive risk factors; and to evaluate the diagnostic performance of the used assays. Subjects and Methods: Fecal specimens were collected from 102 immunocompromised diarrheic children, with ages ranging from few months old to 16 years. All fecal samples were examined microscopically for detection of parasites, as well as immunologically and molecularly for detection of H. pylori and Cryptosporidium spp. Copro-antigens of Cryptosporidium and H. pylori were detected immunologically using rapid chromatographic copro-immunoassay tests. Amplification of H. pylori and Cryptosporidium copro-DNA was performed using the nPCR assay targeting genes encoding H. pylori urease A and Cryptosporidium oocysts wall protein (COWP). Amplified Cryptosporidium PCR products were digested by a restrictive enzyme to detect genotype. Results: H. pylori copro-DNA and copro-antigen were detected in 56 (54.9%) and 18 (17.6%) patients, respectively. Cryptosporidium copro-DNA, and copro-antigen were detected in 22 (21.6%), and 16 (15.8%) patients, respectively, while microscopy detected Cryptosporidium oocysts in only 6 patients (5.9%), with a clear predominance of anthroponotic C. hominis (81%). Cryptosporidium spp. and H. pylori co-infection occurred in 15.8% of patients. None of the studied variables had a significant association with any of the tested pathogens, neither separately nor combined. Conclusion:There was a high detection rate of H. pylori and Cryptosporidium spp. and their co-existence in diarrheic immunocompromised children. Our study results highlight that PCR increased the sensitivity for the diagnosis of Cryptosporidium spp. and H. pylori. More research is needed to establish their relevance.

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“…Sensitivity, specificity, positive predictive value, negative predictive value, accuracy, and kappa agreement were calculated to test the diagnostic yield of the 3 used tests in relation to nPCR. The multiattribute utility theory and analytical hierarchy process were both used to evaluate the lab utility and the diagnostic performance of the used diagnostic procedure according to [ 11 ].…”

Section: Methodsmentioning

confidence: 99%

Multiattribute Analysis of Trichomonas vaginalis Diagnostics and Its Correlation with Clinical Complaints and Contraceptive Methods in a Symptomatic Egyptian Cohort

Ghallab

Alaa

Morsy

2021

Infectious Diseases in Obstetrics and Gynecology

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Background. Trichomonas vaginalis (T. vaginalis) infection has been long considered among the sexually transmitted diseases that possesses a clear effect on women’s health especially in the childbearing period. Methods. A 234 females of age range 18-45 years old attending the Gynecology and Obstetrics Outpatient Clinic of Kasr El Aini Hospitals were enrolled in a cross-sectional study. The taken vaginal swabs were subjected to wet mount microscopy, Giemsa stain, modified Diamond’s culture, and nested polymerase chain reaction (nPCR) amplification. Multiattribute and analytical hierarchy processes were conducted to detect laboratory utility. Univariate and multivariate analyses were done to detect the multiple risk factors that may be associated with Trichomonas infection. Results. Based on nPCR, the prevalence of trichomoniasis was 26.9%. Wet mount, Giemsa stain, and culture showed 100% specificity but of low sensitivity (28.57%, 28.57%, and 57.14%, respectively). On the multivariate analysis, nPCR showed the highest rank for diagnostic performance and culture had the lowest rank. For univariate analysis, there was a significant correlation between T. vaginalis infection and vaginal discharge, burning sensation, dyspareunia, and the use of intrauterine device (IUD) ( P value < 0.05). Conclusion. The routine screening of trichomoniasis using nPCR was reliable, sensitive, and specific. Also, it could financially be considered a more suitable option in batch screening. Significant higher rates of infection were reported among IUD users compared to condom or hormonal-based methods.

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Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt

Cited by 15 publications

References 24 publications

High-Resolution Melting Curve (Hrm) Analysis in Genotypic Discrimination of Cryptosporidium Isolates From Stool of Egyptian Children

High-Resolution Melting Curve (Hrm) Analysis in Genotypic Discrimination of Cryptosporidium Isolates From Stool of Egyptian Children

Cryptosporidium spp. and Helicobacter pylori in a hospitalbased study of diarrheic immunocompromised Egyptian children: Insight into risk factors, and co-infection

Multiattribute Analysis of Trichomonas vaginalis Diagnostics and Its Correlation with Clinical Complaints and Contraceptive Methods in a Symptomatic Egyptian Cohort

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