Evaluation of Ziehl-Neelsen staining, auramine phenol staining, antigen detection enzyme linked immunosorbent assay and polymerase chain reaction, for the diagnosis of intestinal cryptosporidiosis

Abstract

Background and Objectives:Cryptosporidiosis is a very important opportunistic infection and is responsible for significant morbidity and mortality in HIV/AIDS patients. The objective of this study is to evaluate Ziehl-Neelsen staining, auramine phenol staining, antigen detection enzyme linked immunosorbent assay and polymerase chain reaction, for the diagnosis of intestinal cryptosporidiosis.Materials and Methods:The study was designed to determine the efficacy of modified Ziehl-Neelsen (ZN), Auramine-Phenol (AP) staining, antigen detection enzyme linked immunosorbent assay (ELISA) and nested polymerase chain reaction (PCR) for detection of cryptosporidia in 671 HIV-seropositive patients, 353 HIV-seronegative patients including 198 children with diarrhea and 50 apparently healthy adults.Results:Cryptosporidium was detected in 26 (3.9%), 37 (5.5%), 32 (4.8%) and 40 (6%) HIV-seropositive and 8 (2.3%), 10 (2.9%), 9 (2.6%) and 9 (2.6%) HIV-seronegative patients by ZN staining, AP staining, antigen detection ELISA and PCR, respectively. None of the healthy controls were infected with Cryptosporidium. Based on criteria of ‘true positive’ samples, i.e. positive by any two of the four techniques out of ZN, AP, antigen detection ELISA and PCR, sensitivity of ZN and ELISA was 79.06% and 95.35% respectively. AP and PCR were found to be 100% sensitive. Specificity of ZN and ELISA was 100% while specificity of AP and PCR was 99.59% and 99.39% respectively.Conclusions:Auramine phenol staining is a rapid, sensitive and specific technique for diagnosis of intestinal cryptosporidiosis.

Molecular Evaluation of Conventional Microscopic Method Versus Fecal Antigen Capture Enzyme-Linked Immunosorbent Assay and Rapid Immunochromatographic Assay for Diagnosis of Cryptosporidium Infection

“…7,30,[37][38][39] The current study also revealed an apparent higher rate of infection among suspected groups (diarrheic children and immunesuppressed) as compared with apparently healthy ones. The rates of infection detected by different assays are generally within the range (1%-49.1%) previously recorded for Cryptosporidium infection in Saudi Arabia and its neighboring countries.…”

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

“…Using PCR, different Cryptosporidium prevalence rates were reported in Egypt (Hassan et al 2002;El-Mohamady et al 2006;El-Shazly et al 2007;El-Hamshary et al 2008;El-Settawy and Fathy 2012) and worldwide (Morgan et al 1998;Khurana et al 2012;Salyer et al 2012), in Egypt the prevalence ranged from 10.7 to 25.5 %. The difference in the reported prevalence may be attributed to differences in study population, demographic, behavioral, environmental and socioeconomic factors, diagnostic methods, time of the study (summer vs. winter), nutritional status of the children, and educational level of parents (AL-Hindi et al 2007).…”

A multiattribute utility evaluation for the detection of Cryptosporidium species in stool

“…Some authors have advocated the use of negative staining for identification and isolation of C. parvum species. They have also illustrated that the quality of negative staining is time dependent and if the samples are not viewed within a specified time then the spores might become less visible [15]. Moreover, the technique requires higher level of technical and optical skill while using the microscope and owing to this the results might vary among persons with different experience and microscopic skills.…”

“…C. parvum is an important cause of diarrhoea with prevalence rates varying from 1 to 4% in the developed world and 6 to 19% in the developing world as shown in Table 3 [10][11][12][13][14][15]. Among direct microscopy methods -direct wet mount showed a positivity of 22/52 (42.3%), direct staining examination by negative staining showed a positivity of 14/52 (26.9%) while direct staining examination by MZN showed a positivity of 39/52 (75%).…”