Molecular Genotyping of Cryptosporidium spp. and Giardia spp. in Urban Rodents from Kuala Lumpur, Malaysia

Ng, Wern Hann (2017) Molecular Genotyping of Cryptosporidium spp. and Giardia spp. in Urban Rodents from Kuala Lumpur, Malaysia. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Cryptosporidium spp. and Giardia spp. are currently one of the leading causes of diarrhoea globally. There’s currently a lack of information available genetically regarding these two species in Malaysia as there has yet to be a study on the molecular detection of Cryptosporidium and Giardia in rodents in Malaysia. Therefore the main objective of this study is to detect these parasites in urban rodents captured from Kuala Lumpur by employing the polymerase chain reaction (PCR)-based methods and to determine its sub/genotype using phylogenetic analysis. A total of 137 rat samples were captured mostly from wet markets in Sentul and Chow Kit, Kuala Lumpur, Malaysia by Kuala Lumpur City Hall workers from June 2011 until end of July 2011. DNA samples were extracted from the faeces using MN DNA Stool Kit based on protocols of Genomic DNA from soil. Nested-PCR targeting Cryptosporidium spp. and Giardia spp. was performed on the DNA samples. Gel electrophoresis revealed that 24 samples (17.5%) were positive for Cryptosporidium and 4 (2.9%) for Giardia. 20 out of the 24 positive Cryptosporidium spp. were successfully sequenced while 1 out of the 4 positive samples was successfully sequenced for Giardia spp. Phylogenetic analysis confirmed that all positive samples targeted for Cryptosporidium/Giardia spp. were Cryptosporidium parvum subtype IIa and Giardia duodenalis assemblage B.