{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T17:25:06Z","timestamp":1763745906719,"version":"3.45.0"},"reference-count":17,"publisher":"BMJ","issue":"4","content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Clin Pathol"],"accepted":{"date-parts":[[2015,9,14]]},"published-print":{"date-parts":[[2016,4]]},"abstract":"\n Aims<\/jats:title>\n \n The high HIV\/AIDS burden in Sub-Saharan Africa has led to cryptococcosis becoming a public health concern. In this resource-limited setting, conventional identification methods are mainly used to diagnose cryptococcal infections. However, these methods are often inconsistent, and importantly, cannot discriminate between the aetiological agents,\n Cryptococcus neoformans<\/jats:italic>\n and\n C. gattii<\/jats:italic>\n . Therefore, there is a need for an alternative reliable method to identify these species.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n We examined the usefulness of a PCR method, including a restriction digest, in identifying clinical\n C. neoformans<\/jats:italic>\n and\n C. gattii<\/jats:italic>\n isolates. In addition, matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-ToF MS) was performed for validation purposes.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The intraspecific variation between tested strains allowed for their delineation into three traditional varieties of\n C. neoformans<\/jats:italic>\n , that is, varietal forms:\n neoformans<\/jats:italic>\n ,\n grubii<\/jats:italic>\n and\n gattii<\/jats:italic>\n . Furthermore, we uncovered a restriction site (signature sequence: 5\u2032-AATATT-3\u2032) that is present only in the distinct species\n C. neoformans<\/jats:italic>\n (varietal forms\n neoformans<\/jats:italic>\n and\n grubii<\/jats:italic>\n ), and is, importantly, absent in the distinct species\n C. gattii<\/jats:italic>\n (\n C. neoformans<\/jats:italic>\n var.\n gattii<\/jats:italic>\n ). Thus, we were able to discriminate the distinct species by directly digesting the PCR amplicons using the endonuclease\n Ssp<\/jats:italic>\n I. It was also possible to delineate some tested isolates as either\n C. neoformans<\/jats:italic>\n or\n C. gattii<\/jats:italic>\n using our MALDI-ToF MS data.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n The possibility of performing only a restriction digest makes the outlined method, similar to conventional techniques, economical and easy to optimise for routine use in resource-limited settings.<\/jats:p>\n <\/jats:sec>","DOI":"10.1136\/jclinpath-2014-202790","type":"journal-article","created":{"date-parts":[[2015,11,23]],"date-time":"2015-11-23T21:59:00Z","timestamp":1448315940000},"page":"352-357","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":7,"title":["Method for identification of\n Cryptococcus neoformans<\/i>\n and\n Cryptococcus gattii<\/i>\n useful in resource-limited settings"],"prefix":"10.1136","volume":"69","author":[{"given":"Adepemi O","family":"Ogundeji","sequence":"first","affiliation":[{"name":"Department of Microbial, Biochemical and Food Biotechnology, University of the Free State, Bloemfontein, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacobus","family":"Albertyn","sequence":"additional","affiliation":[{"name":"Department of Microbial, Biochemical and Food Biotechnology, University of the Free State, Bloemfontein, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina H","family":"Pohl","sequence":"additional","affiliation":[{"name":"Department of Microbial, Biochemical and Food Biotechnology, University of the Free State, Bloemfontein, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olihile M","family":"Sebolai","sequence":"additional","affiliation":[{"name":"Department of Microbial, Biochemical and Food Biotechnology, University of the Free State, Bloemfontein, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"239","published-online":{"date-parts":[[2015,11,23]]},"reference":[{"key":"2025112109205201000_69.4.352.1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1567-1364.2006.00113.x"},{"key":"2025112109205201000_69.4.352.2","doi-asserted-by":"crossref","first-page":"18466","DOI":"10.7448\/IAS.16.1.18466","article-title":"Disclosure of HIV status to children in resource-limited settings: a systematic review","volume":"16","author":"Vreeman","year":"2013","journal-title":"J Int AIDS Soc"},{"key":"2025112109205201000_69.4.352.3","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0507.2007.01357.x"},{"key":"2025112109205201000_69.4.352.4","first-page":"483","article-title":"Evaluation of conventional and serological methods for rapid diagnosis of cryptococcosis","volume":"127","author":"Saha","year":"2008","journal-title":"Indian J Med Res"},{"key":"2025112109205201000_69.4.352.5","doi-asserted-by":"publisher","DOI":"10.3109\/13693786.2011.602126"},{"key":"2025112109205201000_69.4.352.6","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01271-10"},{"key":"2025112109205201000_69.4.352.7","doi-asserted-by":"publisher","DOI":"10.1111\/j.1567-1364.2002.tb00117.x"},{"key":"2025112109205201000_69.4.352.8","doi-asserted-by":"publisher","DOI":"10.1007\/s11684-010-0011-z"},{"key":"2025112109205201000_69.4.352.9","doi-asserted-by":"publisher","DOI":"10.1111\/j.1567-1364.2006.00088.x"},{"key":"2025112109205201000_69.4.352.10","doi-asserted-by":"publisher","DOI":"10.1139\/W10-030"},{"issue":"(Pt 5)","key":"2025112109205201000_69.4.352.11","first-page":"1209","article-title":"Cryptococcus cyanovorans sp. nov., a basidiomycetous yeast isolated from cyanide contaminated soil","volume":"62","author":"Motaung","year":"2012","journal-title":"Int J Syst Evol Microbiol"},{"key":"2025112109205201000_69.4.352.12","doi-asserted-by":"crossref","unstructured":"White TJ , Bruns T , Lee S , et al . Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA , Gelfand DH , Sninsky JJ , et al , eds. PCR protocols: a guide to methods and applications. San Diego, CA: Academic Press, 1990:315\u201322.","DOI":"10.1016\/B978-0-12-372180-8.50042-1"},{"key":"2025112109205201000_69.4.352.13","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01187-06"},{"key":"2025112109205201000_69.4.352.14","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00651-11"},{"key":"2025112109205201000_69.4.352.15","doi-asserted-by":"publisher","DOI":"10.1080\/mmy.40.1.87.109"},{"key":"2025112109205201000_69.4.352.16","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.2000.0324"},{"key":"2025112109205201000_69.4.352.17","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cir613"}],"container-title":["Journal of Clinical Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jclinpath-2014-202790","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T17:21:05Z","timestamp":1763745665000},"score":1,"resource":{"primary":{"URL":"https:\/\/jcp.bmj.com\/lookup\/doi\/10.1136\/jclinpath-2014-202790"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,11,23]]},"references-count":17,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2016,3,17]]},"published-print":{"date-parts":[[2016,4]]}},"alternative-id":["10.1136\/jclinpath-2014-202790"],"URL":"https:\/\/doi.org\/10.1136\/jclinpath-2014-202790","relation":{},"ISSN":["0021-9746","1472-4146"],"issn-type":[{"type":"print","value":"0021-9746"},{"type":"electronic","value":"1472-4146"}],"subject":[],"published":{"date-parts":[[2015,11,23]]}}}