{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T08:51:42Z","timestamp":1785487902990,"version":"3.56.0"},"reference-count":49,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2019,1,21]],"date-time":"2019-01-21T00:00:00Z","timestamp":1548028800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2019,3]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Meningitis and encephalitis (ME) are central nervous system (CNS) infections mainly caused by bacteria, mycobacteria, fungi, viruses, and parasites that result in high morbidity and mortality. The early, accurate diagnosis of pathogens in the cerebrospinal fluid (CSF) and timely medication are associated with better prognosis. Conventional methods, such as culture, microscopic examination, serological detection, CSF routine analysis, and radiological findings, either are time\u2010consuming or lack sensitivity and specificity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>To address these clinical needs, we developed an advanced fragment analysis (AFA)\u2010based assay for the multiplex detection of 22 common ME pathogens, including eight viruses, 11 bacteria, and three fungi. The detection sensitivity of each target was evaluated with a recombinant plasmid. The limits of detection of the 22 pathogens ranged from 15 to 120\u00a0copies\/reaction. We performed a retrospective study to analyze the pathogens from the CSF specimens of 170 clinically diagnosed ME patients using an AFA\u2010based assay and compared the results with culture (bacteria and fungi), microscopic examination (fungi), polymerase chain reaction (PCR) (<jats:italic>Mycobacterium tuberculosis<\/jats:italic>), and Sanger sequencing (virus) results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The sensitivity of the AFA assay was 100% for 10 analytes. For <jats:italic>Cryptococcus neoformans<\/jats:italic>, the sensitivity was 63.6%. The overall specificity was 98.2%. The turnaround time was reduced to 4\u20106\u00a0hours from the 3\u20107\u00a0days required using conventional methods.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>In conclusion, the AFA\u2010based assay provides a rapid, sensitive, and accurate method for pathogen detection from CSF samples.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcla.22707","type":"journal-article","created":{"date-parts":[[2019,1,22]],"date-time":"2019-01-22T10:39:24Z","timestamp":1548153564000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Development and validation of an advanced fragment analysis\u2010based assay for the detection of 22 pathogens in the cerebrospinal fluid of patients with meningitis and encephalitis"],"prefix":"10.1002","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7429-1996","authenticated-orcid":false,"given":"Fang","family":"Long","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, West China Hospital Sichuan University Chengdu China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mimi","family":"Kong","sequence":"additional","affiliation":[{"name":"Ningbo HEALTH Gene Technologies Co., Ltd. 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