Development of a lateral flow recombinase polymerase amplification assay for rapid and visual detection of Cryptococcus neoformans/C. gattii in cerebral spinal fluid.
Background: For definitive diagnosis of cryptococcal meningitis, Cryptococcus neoformans and/or C. gattii must be identified within cerebral spinal fluid from the patients. The traditional methods for detecting Cryptococcus spp. such as India ink staining and culture are not ideal. Although sensitiv...
| Publicado en: | BMC Infectious Diseases Vol. 19; no. 1; pp. 1 - 10 |
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| Autores principales: | , , , , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
BioMed Central
2/4/2019
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=134549354&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 134549354 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 14712334 1CHU jtl: BMC Infectious Diseases issn: 14712334 maglogo: N pubinfo: dt: 2/4/2019 vid: 19 iid: 1 pid: 24147 pub: BioMed Central artinfo: ui: 134549354 134549354 NLM30717679 134549354 10.1186/s12879-019-3744-6 NLM30717679 134549354 ppf: 1 ppct: 9 formats: tig: atl: Development of a lateral flow recombinase polymerase amplification assay for rapid and visual detection of Cryptococcus neoformans/C. gattii in cerebral spinal fluid. aug: au: Ma, Qinglin Yao, Jilong Yuan, Shixin Liu, Houming Wei, Ning Zhang, Jianming Shan, Wanshui affil: Institute of Maternity and Child Medical Research, Shenzhen Maternity and Child Healthcare Hospital, Southern Medical University, No.2004 Hongli Road, 518028, Shenzhen, Guangdong, China sug: subj: Cryptococcus Cryptococcosis Diagnosis Meningitis, Cryptococcal Diagnosis Polymerase Chain Reaction Methods RNA Cerebrospinal Fluid Microfluidic Analytical Techniques Methods Enzymes Antigens, Fungal DNA Probes Cryptococcosis Cerebrospinal Fluid Temperature Antigens, Fungal Cerebrospinal Fluid RNA Analysis Sensitivity and Specificity Meningitis, Cryptococcal Cerebrospinal Fluid Human Early Diagnosis Validation Studies Comparative Studies Evaluation Research Multicenter Studies Funding Source ab: Background: For definitive diagnosis of cryptococcal meningitis, Cryptococcus neoformans and/or C. gattii must be identified within cerebral spinal fluid from the patients. The traditional methods for detecting Cryptococcus spp. such as India ink staining and culture are not ideal. Although sensitive and specific enough, detection of cryptococcal antigen polysaccharide has a high dose hook effect. Therefore, the aim of this study was to introduce a new rapid and simple detection method of Cryptococcus neoformans and C. gattii in cerebral spinal fluid.Methods: The lateral flow strips combined with recombinase polymerase amplification (LF-RPA) assay was constructed to detect the specific DNA sequences of C. neoformans and C. gattii. The detection limit was evaluated using serial dilutions of C. neoformans and C. gattii genomic DNA. The specificity was assessed by excessive amount of other pathogens genomic DNA. The optimal detection time and amplification temperature were also analyzed. The diagnostic parameters were first calculated using 114 clinical specimens and then compared with that of other diagnostic method. A brief analysis and comparison of different DNA extraction methods was discussed, too.Results: The LF-RPA assay could detect 0.64 pg of genomic DNA of C. neoformans per reaction within 10 min and was highly specific for Cryptococcus spp.. The system could work well at a wide range of temperature from 25 to 45 °C. The overall sensitivity and specificity were 95.2 and 95.8% respectively. As amplification template for LF-RPA assay, both cell lysates and genomic DNA produce similar experimental results.Conclusions: The LF-RPA system described here is shown to be a sensitive and specific method for the visible, rapid, and accurate detection of Cryptococcus spp. in cerebral spinal fluid and might be useful for clinical preliminary screening of cryptococcal meningitis. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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