The Prevalence of Cryptosporidium among Children Hospitalized because of Gastrointestinal Symptoms and the Efficiency of Diagnostic Methods for Cryptosporidium

Abstract

Cryptosporidium is a parasite that causes watery diarrhea among both children and adults. However, because many physicians do not routinely ask for Cryptosporidium diagnostic test, cryptosporidiosis prevalence is likely underestimated. The current study investigated the prevalence of cryptosporidiosis among children admitted to the hospital with gastrointestinal symptoms. Stool sample was collected from each child and subjected to routine microbiological culture. Cryptosporidium presence was tested by three different methods: real-time PCR (RT-PCR), quick antigen, and microscopic examination with acid-fast staining. Each positive specimen was further tested with BioFire FilmArray Multiplex PCR (bioMérieux SA, Marcy-l'Etoile, France) to determine that Cryptosporidium is the only pathogen in the sample. Demographic and epidemiological data were collected from the patients' medical records. Of 291 patient stool samples, nine were positive for Cryptosporidium hominis or Cryptosporidium parvum. The average age of the nine Cryptosporidium-positive cases was 2.3 years, lower than the average age of the study population (4.2 years). Of the positive cases, 66.7% were Arabs and 33.3% were Jews. The main complaint of children with Cryptosporidium was diarrhea. Regarding the laboratory methods for Cryptosporidium identification, of the positive samples, 100% (9/9) were identified by RT-PCR, 88% (8/9) were positive by antigen test, and only 67% (6/9) were positive by microscopic examination with acid-fast staining. Because of the low incidence of Cryptosporidium among patients in our facility, the prevalence of cryptosporidiosis still cannot be established. Nevertheless, among the other pathogens found in stool samples, Cryptosporidium is the second leading cause of hospitalization due to gastrointestinal disease in children in our area.

Protozoan infections are under-recognized in Swedish patients with gastrointestinal symptoms

“…In this study, there was no statistically significant difference in detection between microscopy and real-time PCR in the two groups in which all samples were tested with both methods. In other studies, real-time PCR has increased detection compared to microscopy [ 32 – 34 ].…”

Prevalence of Cryptosporidium spp. in Yaks (Bos grunniens) in China: A Systematic Review and Meta-Analysis

“…PCR can be used to detect complete DNA and fragments of parasites, and has become the best method for detecting Cryptosporidium spp. ( Efrat et al., 2019 ). Thus, we suggest that the researchers to use the PCR method for detecting Cryptosporidium spp.…”

Development of duplex real time PCR for quick detection of Cryptosporidia in goats

“…The 18ssu rRNA gene is highly abundant in terms of genomic copies and it has high transcripts that can increase the sensitivity of the assay, while the COWP is available only in transcriptionally active live cryptosporidial oocysts. (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(28)(29)(30)(31) Hence, in the current study we report an mRNA based TaqMan® probe duplex real time PCR that can able to detect the metabolically active Cryptosporidium oocysts from dead oocysts in passively infected animals…”

“…Microscopy is an excellent tool and can be highly specific but requires expertise and experience in identification of the weak acid fast round oocysts from other artefacts similar to them. Besides conventional microscopy and staining, molecular diagnostic approaches like PCR are also gaining momentum Molecular diagnostic assays are usually regarded as more sensitive compared to microscopy and serological diagnosis in human and animals for the detection of Cryptosporidium in several studies (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). The molecular assays are determined by the factors like stage of life cycle/infection as well as method of nucleic acid extraction for targeting the Cryptosporidium spp.…”