Enhanced Commendable Sensitivity and Specificity for MSI in Colorectal Cancer by a New PCR-HRM Based 8-Loci MSI Assay

IF 2.6 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Clinical Laboratory Analysis Pub Date : 2024-08-12 DOI:10.1002/jcla.25085

Xueping Xiang, Xiaojing Ma, Linlin Ying, Hong Zou

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Abstract

Background

This study evaluated the performance of the PCR-HRM assay by comparing it with immunohistochemistry (IHC) for mismatch repair (MMR) proteins and the PCR capillary electrophoresis (PCR-CE) methods.

Results

A total of 224 patients with colorectal cancer participated in the study, with nearly half having mismatch repair deficiency (dMMR) tissues and the remainder possessing pMMR tissues. There was a 97.77% concordance between the PCR-HRM assay and IHC, and a 97.56% concordance between PCR-HRM and the PCR-CE assay. In comparison with IHC for dMMR proteins, the PCR-HRM demonstrated a sensitivity of 96.36% and a specificity of 99.12%. When juxtaposed with the PCR-CE assay, its sensitivity was 98.96% and specificity stood at 96.33%. The mutations observed in the microsatellite loci were uniformly distributed across all eight loci. Discrepant outcomes were more frequent in instances of MLH1 and PMS2 deficiency. Furthermore, the germline mutation status of MLH1, MSH2, PMS2, and MSH6 in 62 patients was ascertained using next-generation sequencing. All patients displaying MMR gene pathogenic mutations (N = 14) were identified as MSI-H by PCR-HRM, whereas those with MSS tissues (N = 43) did not exhibit MMR gene pathogenic mutations. Thus, the PCR-HRM method proficiently pinpoints tumors with verified germline MMR mutations, indicative of Lynch syndrome.

Conclusion

Conclusively, the PCR-HRM assay emerges as a swift and congruent diagnostic tool for microsatellite instability, boasting commendable sensitivity and specificity in colorectal cancer.

Abstract Image

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基于 PCR-HRM 的新型 8 个基因 MSI 检测法提高了结直肠癌 MSI 检测的灵敏度和特异性。

背景：这项研究通过比较PCR-HRM检测法与错配修复（MMR）蛋白免疫组化（IHC）法和PCR毛细管电泳（PCR-CE）法，评估了PCR-HRM检测法的性能：共有 224 名结直肠癌患者参与了研究，其中近一半患者的组织存在错配修复缺陷（dMMR），其余患者的组织存在 pMMR。PCR-HRM 检测与 IHC 检测的一致性为 97.77%，PCR-HRM 检测与 PCR-CE 检测的一致性为 97.56%。与 IHC 检测 dMMR 蛋白相比，PCR-HRM 的灵敏度为 96.36%，特异性为 99.12%。与 PCR-CE 检测法相比，其灵敏度为 98.96%，特异性为 96.33%。在微卫星位点上观察到的突变均匀分布在所有八个位点上。在 MLH1 和 PMS2 缺乏的情况下，异常结果更为常见。此外，62 名患者的 MLH1、MSH2、PMS2 和 MSH6 的种系突变状态也通过下一代测序得以确定。通过 PCR-HRM，所有显示 MMR 基因致病突变的患者（14 人）都被确定为 MSI-H，而那些有 MSS 组织的患者（43 人）则没有显示 MMR 基因致病突变。因此，PCR-HRM方法能有效地确定肿瘤是否存在已验证的种系MMR基因突变，这也是林奇综合征的指征：最后，PCR-HRM 检测是一种快速、一致的微卫星不稳定性诊断工具，在结直肠癌中具有值得称道的灵敏度和特异性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Clinical Laboratory Analysis 医学-医学实验技术

CiteScore

5.60

自引率

7.40%

发文量

584

审稿时长

6-12 weeks

期刊介绍： Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.