{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T03:33:32Z","timestamp":1774668812840,"version":"3.50.1"},"reference-count":19,"publisher":"BMJ","issue":"7","funder":[{"name":"Hui Ming"},{"name":"Michael Seak-Kan Tong"},{"name":"Dr Richard Yu and Carol Yu"},{"name":"Hui Hoy"},{"name":"Shaw Foundation Hong Kong"},{"name":"High Level Hospital-Summit Program in Guangdong, China"},{"name":"Chan Yin Chuen Memorial Charitable Foundation"},{"name":"Chow Sin Lan Charity Fund Limited"}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Clin Pathol"],"accepted":{"date-parts":[[2019,3,19]]},"published-print":{"date-parts":[[2019,7]]},"abstract":"<jats:sec>\n                  <jats:title>Aims<\/jats:title>\n                  <jats:p>Helminth infections are becoming uncommon in high-income countries and laboratory staff may lose expertise in their morphological identification, especially in histological sections where speciation of helminths is challenging. Commercially available molecular diagnostic panels for faecal specimens only offer tests for protozoa but not helminths. We aim to improve the identification accuracy of helminths using a multiplex PCR assay.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We designed three pairs of PCR primers and probes targeting multicopy genes for a multiplex single-tube real-time PCR assay which covers 16 trematode (28S rRNA gene), 24 cestode (<jats:italic>cox1<\/jats:italic> gene) and 33 nematode (<jats:italic>cox1<\/jats:italic> gene) species. Helminths (n=27) from faecal samples (n=10), fresh parasites (n=11), formalin-fixed specimens (n=4), cerebrospinal fluid (n=1) and bile (n=1) were examined morphologically and tested by PCR. Fifty stool samples negative for parasites by microscopy were also tested.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>The PCR assay correctly identified the genera of all tested helminths. Agarose gel electrophoresis and sequencing of the purified PCR amplicons confirmed that the PCR products were of correct sizes with 100% correlation with the respective species. Sequencing of the <jats:italic>cox1<\/jats:italic> gene failed to identify <jats:italic>Capillaria<\/jats:italic> spp. in one sample owing to the lack of corresponding sequences in GenBank. PCR and sequencing of the nematode 18S rRNA gene using consensus primers showed 100% homology with <jats:italic>Capillaria<\/jats:italic> spp. sequence. No positive PCR products were found in the negative stool samples.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The highly specific test correctly identified all helminths in our cohort. It is a useful adjunct to helminth identification in difficult situations such as histological sections.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jclinpath-2018-205590","type":"journal-article","created":{"date-parts":[[2019,4,5]],"date-time":"2019-04-05T17:15:54Z","timestamp":1554484554000},"page":"487-492","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":6,"title":["Improving the specific diagnosis of trematode, cestode and nematode infections by a multiplex single-tube real-time PCR assay"],"prefix":"10.1136","volume":"72","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0266-4401","authenticated-orcid":false,"given":"Samson S Y","family":"Wong","sequence":"first","affiliation":[{"name":"Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China"},{"name":"Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosana W S","family":"Poon","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Queen Mary Hospital, Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kelvin K W","family":"To","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China"},{"name":"Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China"},{"name":"Department of Clinical Microbiology and Infection Control, The University of Hong Kong\u2013Shenzhen Hospital, Shenzhen, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jasper F W","family":"Chan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China"},{"name":"Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China"},{"name":"Department of Clinical Microbiology and Infection Control, The University of Hong Kong\u2013Shenzhen Hospital, Shenzhen, China"},{"name":"Hainan Medical University\u2013The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, and The University of Hong Kong, Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gang","family":"Lu","sequence":"additional","affiliation":[{"name":"Hainan Medical University\u2013The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, and The University of Hong Kong, Hong Kong, China"},{"name":"Department of Pathogen Biology, Hainan Medical University, Haikou, Hainan, China"},{"name":"Key Laboratory of Translational Tropical Medicine, Hainan Medical University, Haikou, Hainan, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fanfan","family":"Xing","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology and Infection Control, The University of Hong Kong\u2013Shenzhen Hospital, Shenzhen, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vincent C C","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Queen Mary Hospital, Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kwok-Yung","family":"Yuen","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China"},{"name":"Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China"},{"name":"Department of Microbiology, Queen Mary Hospital, Hong Kong, China"},{"name":"Department of Clinical Microbiology and Infection Control, The University of Hong Kong\u2013Shenzhen Hospital, Shenzhen, China"},{"name":"Hainan Medical University\u2013The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, and The University of Hong Kong, Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"239","published-online":{"date-parts":[[2019,4,5]]},"reference":[{"key":"2025092610055644000_72.7.487.1","doi-asserted-by":"publisher","DOI":"10.1177\/1535370214523880"},{"key":"2025092610055644000_72.7.487.2","doi-asserted-by":"crossref","unstructured":"Garcia LS . Diagnostic medical parasitology. 6th edition. 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Diagnostic pathology of parasitic infections with clinical correlations. Second edition. 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