Detection of Cryptosporidium sp infection by PCR and modifi ed acid fast staining from potassium dichromate preserved stool.
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Date
2009-07
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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 modifi ed 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 modifi ed 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 signifi cant 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.
Description
Keywords
18S rRNA, cryptosporidiosis
Citation
Kurniawan Agnes, Dwintasari Sri W, Soetomenggolo Herbowo A, Wanandi Septelia I. Detection of Cryptosporidium sp infection by PCR and modifi ed acid fast staining from potassium dichromate preserved stool. Medical Journal of Indonesia. 2009 Jul; 18(3): 147-152.