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All Journal Medical Journal of Indonesia Paediatrica Indonesiana
Herbowo A. Soetomenggolo
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Health Professions Medicine & Pharmacology

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Detection of Cryptosporidium sp infection by PCR and modified acid fast staining from potassium dichromate preserved stool Kurniawan, Agnes; Dwintasari, Sri W.; Soetomenggolo, Herbowo A.; Wanandi, Septelia I.
Medical Journal of Indonesia Vol 18, No 3 (2009): July-September
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (421.766 KB) | DOI: 10.13181/mji.v18i3.354

Abstract

Aim To identify the frequency of Cryptosporidium infection in children below 3 years old by examining concentrated long term preserved stool using PCR detection of 18S rRNA gene and compared with modified acid fast staining technique.Methods Hundred eighty eight stools from children ≤ 3 years old were stored for 13 months in 2.5% K2Cr2O7 solution at 40C. Cryptosporidium oocysts were isolated by water-ether concentration technique. The concentrates were smeared onto object glass and stained with modified acid fast staining, and the rest of the concentrates were DNA extracted by freezing and thawing cycles and proteinase K digestion, then direct PCR was done to detect 18S rRNA gene.Result The proportion of positive stools for Cryptosporidium sp by acid fast staining from concentrated stools and 18S rRNA PCR were 4.8% and 34.6% respectively, which showed statistically significant difference.Conclusion The frequency of Cryptosporidium infection among children ≤ 3 years old was very high and stool storage in K2Cr2O7 for 13 months did not affect the PCR result. High prevalence of Cryptosporidium infection indicated high transmission in that area and the potential to be transmitted to other individuals such as the immunocompromised. (Med J Indones 2009;18:147-52)Key words: 18S rRNA, cryptosporidiosis
Cryptosporidiosis in children less than three years old in Ciliwung Riverside, Kampung Melayu Village, Jakarta, Indonesia Herbowo A. Soetomenggolo; Agus Firmansyah; Agnes Kurniawan; Partini P. Trihono
Paediatrica Indonesiana Vol 48 No 2 (2008): March 2008
Publisher : Indonesian Pediatric Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (364.612 KB) | DOI: 10.14238/pi48.2.2008.99-103

Abstract

Background Cryptosporidium infection is often found in children,especially children below three years old. Many risk factors canaffect cryptosporidiosis prevalence. At this moment, the prevalenceand risk factors of cryptosporidiosis in children in Jakarta areunknown.Objectives To determine the cryptosporidiosis prevalence, clinicalmanifestations, and risk factors in children below three years old.Methods This cross sectional study involved 474 subjects betweenthe age of 0 to 35 months in Ciliwung riverside, Kampung Melayuvillage, from December 2005 until April 2006. Stool specimenswere examined using modified acid-fast staining. Nutritional statuswas measured based on actual body weight over ideal body weightratio (NCHS-CDC 2000).Results Cryptosporidium cysts were found in stool sample of 10/474 subject (2.1%). Most of the cases used ground water as asource for drinking and washing. All positive cases lived in houseswith bad sanitation, flooded house and 9/10 cases had a crowdedhousehold. Cat and mice were the two most frequently foundanimals around the house. We found five asymptomatic cases andall of the cases were undernourished.Conclusions The prevalence of cryptosporidiosis in this study is2.1%. Due to small number of cases no risk factor could beidentified. Use of groundwater as a water source, bad sanitation,cat and mice around the house, flooded house, crowded householdand undernourishment might be related to cryptosporidiosisprevalence. Half of the infected children were asymptomatic.
Co-Authors Agnes Kurniawan Agus Firmansyah Partini P. Trihono Septelia I. Wanandi Sri W. Dwintasari