{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T01:57:01Z","timestamp":1774663021251,"version":"3.50.1"},"reference-count":28,"publisher":"Microbiology Society","issue":"6","license":[{"start":{"date-parts":[[2023,6,19]],"date-time":"2023-06-19T00:00:00Z","timestamp":1687132800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001277","name":"Drinking Water Inspectorate","doi-asserted-by":"crossref","award":["DWI 70\/2\/332"],"award-info":[{"award-number":["DWI 70\/2\/332"]}],"id":[{"id":"10.13039\/501100001277","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,6,19]]},"abstract":"<jats:p>\n                  <jats:italic>\n                     <jats:named-content content-type=\"#213368\">A corrigendum of this article has been published full details can be found at 10.1099\/jmm.0.001926<\/jats:named-content>\n                  <\/jats:italic>\n               <\/jats:p>\n               <jats:p>\n                  <jats:bold>Introduction.<\/jats:bold>\n                  <jats:italic>Cryptosporidium<\/jats:italic> presents one of the main waterborne public health threats due to its resistance to chlorine disinfection and ability to cause large-scale outbreaks. The standard method used in the UK water industry for detection and enumeration of <jats:italic>Cryptosporidium<\/jats:italic> is based on fluorescence microscopy and is laborious and expensive. Molecular methods such as quantitative polymerase chain reaction (qPCR) can be more amenable to streamlining through automation, improving workflows and standardizing procedures.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Hypothesis.<\/jats:bold> The null hypothesis was that there was no difference in the detection or enumeration between the standard method and a qPCR.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Aim.<\/jats:bold> We aimed to develop and evaluate a qPCR for the detection and enumeration of <jats:italic>Cryptosporidium<\/jats:italic> in drinking water, and to compare the assay with the standard method used in the UK.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Methodology.<\/jats:bold> We first developed and evaluated a qPCR method by incorporating an internal amplification control and calibration curve into a real-time PCR currently used for <jats:italic>Cryptosporidium<\/jats:italic> genotyping. Then we compared the qPCR assay with the standard method of immunofluorescent microscopy for the detection and enumeration of 10 and 100 <jats:italic>Cryptosporidium<\/jats:italic> oocysts in 10 l of artificially contaminated drinking water.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Results.<\/jats:bold> The results demonstrated that detection of <jats:italic>Cryptosporidium<\/jats:italic> by this qPCR was reliable at low numbers of oocysts; however, enumeration was less reliable and more variable than immunofluorescence microscopy.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Conclusions.<\/jats:bold> Despite these results, qPCR offers practical advantages over microscopy. There is potential for the use of PCR-based methods for <jats:italic>Cryptosporidium<\/jats:italic> analysis if parts of the upstream sample preparation are revised, and alternative technologies for enumeration (such as digital PCR) are also explored to improve analytical sensitivity.<\/jats:p>","DOI":"10.1099\/jmm.0.001715","type":"journal-article","created":{"date-parts":[[2023,6,19]],"date-time":"2023-06-19T13:17:24Z","timestamp":1687180644000},"source":"Crossref","is-referenced-by-count":4,"title":["A comparison of qPCR and microscopy for the detection and enumeration of Cryptosporidium oocysts from drinking water"],"prefix":"10.1099","volume":"72","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6594-2408","authenticated-orcid":false,"given":"Guy","family":"Robinson","sequence":"first","affiliation":[{"name":"Swansea University Medical School, Swansea, UK"},{"name":"Cryptosporidium Reference Unit, Public Health Wales, Swansea, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1024-2276","authenticated-orcid":false,"given":"Kristin","family":"Elwin","sequence":"additional","affiliation":[{"name":"Cryptosporidium Reference Unit, Public Health Wales, Swansea, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Jones","sequence":"additional","affiliation":[{"name":"Present address: Wessex Water Operations Centre, Claverton Down, Bath, UK"},{"name":"D\u0175r Cymru Welsh Water, Glaslyn, The Avenue, Coedkernew, Duffryn, Newport, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9309-7096","authenticated-orcid":false,"given":"Rachel M.","family":"Chalmers","sequence":"additional","affiliation":[{"name":"Cryptosporidium Reference Unit, Public Health Wales, Swansea, UK"},{"name":"Swansea University Medical School, Swansea, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2004.09.002"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2021.150562"},{"key":"R3","article-title":"Risk Assessment of Cryptosporidium in Drinking-water. World Health Organization 2009, Geneva"},{"key":"R4","article-title":"Guidelines for drinking-water quality. Fourth edition incorporating the first and second addenda World Health Organization 2022, Geneva"},{"key":"R5","article-title":"ISO 15553:2006 Water quality Isolation and identification of Cryptosporidium oocysts and Giardia cysts from water","year":"2006"},{"key":"R6","article-title":"Method 1623.1: Cryptosporidium and Giardia in Water by Filtration\/IMS\/FA. Office of Water","year":"2012"},{"key":"R7","article-title":"The Microbiology of Drinking Water 2010) - Part 14 - Methods for the isolation, identification and enumeration of Cryptosporidium oocysts and Giardia. 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