Use of Markers to Determine Cryptosporidium Genotypes for Epidemiology Tracking and Detection

Abstract

Polymerase chain reaction (PCR) enables amplification of specific DNA fragments for the detection and tracking of Cryptosporidium spp. Newly obtained DNA are compared to an ever-growing database of Cryptosporidium sequences with uncertain or outdated metadata in primary public repositories (i.e., EMBL/DDBJ/GenBank). Here, we describe standard operating procedures to obtain DNA sequences from Cryptosporidium spp. marker genes. Small-subunit ribosomal RNA gene, large-subunit ribosomal RNA gene, and glycoprotein 60 are amplified using conventional PCR. Amplified and sequenced genes are compared to a reference library of up-to-date curated gene sequences to identify Cryptosporidium species and variants.