Utility of Different Diagnostic Tools for Detection of Cryptosporidium in Children with Diarrhea

Abstract

Background: Cryptosporidiosis is a worldwide zoonotic parasitic disease found in children and HIV-positive individuals, and is mainly accompanied with diarrhea. Objectives: This study attempted to compare the sensitivity and specificity of acid-fast (AF) staining, polymerase chain reaction (PCR), and enzyme-linked immunosorbent assay (ELISA) methods to determine Cryptosporidium and its predominant species in diarrheal stool samples of children. Methods: In total, 221 diarrheal stool samples were collected from children admitted to Motahari Hospital in Urmia city, NorthWest of Iran. The AF staining and ELISA methods were used to analyze all the samples, while the PCR method was considered as the gold standard of the study. Positive samples shown by PCR were sequenced to determine Cryptosporidium species (spp.). The three methods were compared regarding statistical factors, sensitivity, specificity, positive and negative predictive values, time duration, and experiment costs. Results: Of the 221 analyzed samples, four and seven samples were positive for Cryptosporidium, as indicated by the AF staining and PCR methods, respectively. The sensitivity and specificity of the AF staining method were shown to be 57.14% and 99.53%, respectively, and five out of 94 samples were diagnosed as positive using ELISA (with 71.4% sensitivity and 100% specificity). Conclusions: Our findings showed that ELISA found less false positive and false negative in comparison with the AF staining method to detect Cryptosporidium. Despite having higher sensitivity and specificity in comparison with the ELISA method, PCR needs more time and cost; therefore, ELISA is preferred for laboratory routine works. Due to limited information on molecular epidemiology of Cryptosporidium spp., more studies using molecular methods are needed to elucidate accurate species.

Genotyping of Cryptosporidium species in children suffering from diarrhea in Sharkyia Governorate, Egypt

“…Regarding age distribution of infection, the present study showed that Cryptosporidium infection was more frequent among children aged 1-5 years. These results were in agreement with Al-Jawabreh et al [44] and Mahmoudi et al [43], who reported that the highest frequency of cryptosporidiosis was in children under five years. This may be because children in this age group eat without washing their hands, lack knowledge about hygiene and immaturity of gut mucosa [45].…”

“…In the present work, Cryptosporidium infection was more common among children with history of animal contact as they were (66.7%) of the total positive cases with statistically significant difference (p = 0.01). In the same manner, Hossain et al [39] and Mahmoudi et al [43] revealed a positive correlation between Cryptosporidium infection and animal contact. The infected livestock animals (cattle and sheep) have the liability to excrete a large number of Cryptosporidium oocysts, contaminate the environment, and infect individuals who live in close proximity to these animals, particularly in rural areas [47].…”

“…showed a predominance of C. parvum in 92% of infected children. In Iran, Mahmoudi et al [43] found that all isolates of the species from children were identified as C. parvum.…”

Use of Recombinant CP2 and CP23 Antigens of Cryptosporidium parvum for Serodiagnosis of Human Cryptosporidiosis

“…The fecal smear preparation, Ziehl-Neelsen staining, and microscopic observation are routinely used to diagnose cryptosporidiosis; however, these methods have been proved to be ineffectual due to low sensitivity and specificity [ 3 ] . Therefore, molecular tests have been employed for final confirmation of the diagnosis [ 4 ] , though it needs expertise and specialized equipment [ 4 , 5 ] .…”