Genetic characterization of selected parasites from people with histories of gastrointestinal disorders using a mutation scanning‐coupled approach

Abstract

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 DNAs 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, Giardia, and strongylid nematodes were detected in 94, 132 and 12 samples. Cryptosporidium hominis subgenotypes IbA10G2, IdA15G1, IgA17, IgA18, and IfA13G1 were identified in 74.6, 16.9, 5.6, 1.4, and 1.4% of 71 samples, respectively. For Cryptosporidium parvum, subgenotypes IIaA17G2R1 (47.6%) and IIaA18G3R1 (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 Ancylostoma ceylanicum (n = 2), Ancylostoma duodenale (4), Necator 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.

Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments

“…cuniculus , a novel Cryptosporidium species most closely related to C . wrairi and the Cryptosporidium mink genotype have been reported [24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42]. The aim of the present study was to use molecular tools to identify the Cryptosporidium sp.…”

Zoonotic Ancylostoma ceylanicum Infection Detected by Endoscopy

“…However, no clinical information was available. 13 In our case, although the laboratory results showed that hemoglobin was slightly below the normal limit, it may not have been caused by A. ceylanicum infection, because only a single worm was observed and recovered during endoscopy.…”

Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia

“…Although gene microarray is rapid and specific, probe synthesis cycle is longer, and the cost is expensive (Wang et al 2004). Single-strand conformation polymorphism (SSCP) has advantage of lower cost, except complex operation and bad specificity (Koehler et al 2013). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is low requirement for equipment, but with ethidium bromide, and manual operation for every step.…”