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Genetic characterization of selected parasites from people with histories of gastrointestinal disorders using a mutation scanning‐coupled approach
Author(s) -
Koehler Anson V.,
Bradbury Richard S.,
Stevens Melita A.,
Haydon Shane R.,
Jex Aaron R.,
Gasser Robin B.
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300100
Subject(s) - biology , internal transcribed spacer , giardia , genomic dna , giardia lamblia , ribosomal dna , cryptosporidium , ribosomal rna , feces , polymerase chain reaction , genetics , single strand conformation polymorphism , haemonchus contortus , gene , helminths , microbiology and biotechnology , zoology , phylogenetics
A SSCP analysis and targeted sequencing approach was used for the genetic characterization of some major pathogens from a cohort of 227 people with histories of gastrointestinal disorders. Genomic DNA s from fecal samples were subjected to PCR ‐amplification of regions in the glycoprotein ( gp60 ) or triose phosphate isomerase ( tpi ) gene, or the second internal transcribed spacer of nuclear ribosomal DNA ( ITS ‐2). Cryptosporidium , G iardia , and strongylid nematodes were detected in 94, 132 and 12 samples. Cryptosporidium hominis subgenotypes I b A 10 G 2, I d A 15 G 1, I g A 17, I g A 18, and I f A 13 G 1 were identified in 74.6, 16.9, 5.6, 1.4, and 1.4% of 71 samples, respectively. For Cryptosporidium parvum , subgenotypes II a A 17 G 2 R 1 (47.6%) and II a A 18 G 3 R 1 (23.8%) were identified in 23 samples. Giardia duodenalis assemblage B (78%) was more common than assemblage A (22%). In addition, DNA of the nematodes A ncylostoma ceylanicum ( n = 2), A ncylostoma duodenale (4), N ecator americanus (5), and Haemonchus contortus (1) was specifically detected. This is the first report of A. ceylanicum in two persons in Australia and, we provide molecular evidence of H. contortus in a child. This SSCP ‐based approach should provide a useful diagnostic and analytical tool for a wide range of pathogens.