This study employed the post-real-time PCR application, high resolution melting (HRM) analysis, in order to differentiate between characterised clinical and reference Cryptosporidium parvum samples obtained from Cork University Hospital (Cork, Ireland) and the Cryptosporidium Reference Unit (Swansea, Wales). A sample set composed of 18 distinct C. parvum gp60-subtypes of the IIa gp60-subtype family (an allele family accounting for over 80% of all cryptosporidiosis cases in Ireland) was employed. HRM analysis-based interrogation of the gp60, MM5 and MS9-Mallon tandem repeat loci was found to completely differentiate between 10 of the 18 studied gp60-subtypes. The remaining eight gp60-subtypes were differentiated into three distinct groupings, with the designations within these groupings resolved to two to three potential gp60-subtypes.

The current study aimed to develop a novel, reproducible, real-time PCR based multi-locus genotyping method to distinguish between C. parvum gp60-subtypes. These preliminary results support the further expansion of the multi-locus panel in order to increase the discriminatory capabilities of this novel method.

本研究中所用的后实时PCR应用，高分辨率熔解（HRM）分析，以便表征临床和参考之间进行区分小隐孢子虫从科克大学医院（黄柏，爱尔兰）和所得到的试样隐参考单元（斯旺西，威尔士）。使用了由IIa gp60亚型家族（等位基因家族占爱尔兰所有隐孢子虫病病例的80％以上）的18种不同的小隐球菌gp60亚型组成的样本集。发现基于HRM分析的对gp60，MM5和MS9-Mallon串联重复基因座的询问可以完全区分18个研究的gp60亚型中的10个。剩下的八gp60-亚型被分为三个不同的组，这些组中的名称被解析为两到三个潜在的 gp60-亚型。

当前的研究旨在开发一种新颖的，可再现的，基于实时PCR的多基因座基因分型方法，以区分小球藻gp60亚型。这些初步结果支持多场所面板的进一步扩展，以增加这种新颖方法的区分能力。