Routine and outbreak monitoring of Cryptosporidium using a molecular diagnostic method

IF 1.9 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2025-09-17 DOI:10.1016/j.mimet.2025.107271

Mora-Ruiz Merit, Torres-Martin Ana, Cuberos-Gomez Lucila

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引用次数: 0

Abstract

Cryptosporidiosis is a significant public health disease associated with various water matrices. In response to the increase of outbreaks reported in several countries and the limitations of conventional detection methods, this study presents and validates a quantitative molecular protocol based on qPCR for the detection of Cryptosporidium spp. and Cryptosporidium parvum in different water matrices. Our method includes high-volume water filtration, thermal-mechanic-enzymatic cell disruption and an optimized qPCR assay, resulting in a highly sensitive and reproducible method. We evaluated the sensitivity, specificity, turbidity interference, and performance compared to the conventional IMS-IM method. Validation over 27 months demonstrated robust performance, with recovery rates up to 55.1 %, detection of as few as 9 oocysts per sample and a cost reduction of over 35 % compared to traditional methods. The method was successfully applied during recent outbreak investigations and offers a reliable, rapid, and scalable alternative for environmental Cryptosporidium monitoring, with strong potential for integration into regulatory frameworks.

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隐孢子虫分子诊断方法的常规监测和疫情监测。

隐孢子虫病是一种与各种水基质相关的重大公共卫生疾病。针对一些国家报告的隐孢子虫疫情增加和传统检测方法的局限性，本研究提出并验证了基于qPCR的不同水基质中隐孢子虫和细小隐孢子虫的定量分子检测方案。我们的方法包括大容量水过滤，热-机械-酶细胞破坏和优化的qPCR分析，从而产生高灵敏度和可重复性的方法。与传统的IMS-IM方法相比，我们评估了灵敏度、特异性、浊度干扰和性能。27个 月的验证显示了稳健的性能，回收率高达55.1% %，每个样品的检出率低至9个卵囊，与传统方法相比，成本降低了35% %以上。该方法已成功应用于最近的疫情调查，为隐孢子虫环境监测提供了一种可靠、快速和可扩展的替代方法，具有纳入监管框架的巨大潜力。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.