gattii ( Bromilow and Corcoran, 2007 Chen et al., 2008). neoformans is a more common causative agent of cryptococcal infections in China and elsewhere compared to C. gattii can establish Cryptococcus infections and infect the lungs, exhibiting a high pathogenicity that is difficult to treat. neoformans is the main pathogen of cryptococcosis and mainly affects immunocompromised individuals ( Rodrigues et al., 1999 Voelz and May 2010), showing a preference for CNS infection ( Rodrigues et al., 2007 Bielska and May 2016 May et al., 2016). Early diagnosis is important for treatment of this disease ( Shaheen et al., 2018). Thus, the clinical mortality rate of these fungi is extremely high, causing more than 180,000 deaths worldwide each year. Untreated CM has a 100% mortality rate, a 10–40% mortality rate after treatment, and a 20–25% recurrence rate in survivors. IntroductionĬryptococcus neoformans and Cryptococcus gattii are the two main pathogenic cryptococci that cause human infections ( Torda et al., 2001). With the method established in this study, test results are provided rapidly, and through the correct and timely administration of drugs and aggressive treatment by clinicians, the maximum number of critically ill patients can be saved. The traditional culture method can delay the diagnosis of CM and cause irreversible damage due to its time-consuming nature. gattii at 37☌ for 20 min, which allows for rapid and accurate diagnosis of critically ill patients. The RPA-LFS established in this study can detect C. gattii, meeting the need for rapid, specific, and sensitive detection. A total of 487 clinical samples collected from multicenter multiplexes were tested to evaluate the detection performance of the RPA-LFS system, which revealed that the system could specifically detect C. This system was developed to obtain results in 20 min at an isothermal temperature of 37☌ with a lower limit of detection as low as 10 CFU/μL or 1 fg/μL. gattii, and the primer-probe design was optimized by introducing base mismatches to obtain a specific and sensitive primer-probe combination for clinical testing, and specificity of the detection system was determined for 26 common clinical pathogens. In our study, RPA- lateral flow strip (LFS) was used to amplify the capsule-associated gene, CAP64, of C. Recombinase polymerase amplification (RPA) is a promising isothermal amplification technique that can compensate for the shortcomings of the above techniques, featuring short reaction time, high specificity, and high sensitivity, thus meeting the demand for in-field detection of C.neoformans/ C. gattii infection, there is a desperate need for fast, sensitive, and on-site detection methods to meet the clinical diagnosis. Due to the rapid progress of meningitis caused by C. In addition, enzyme-linked immunosorbent assay (ELISAs), polymerase chain reaction (PCR), real-time quantitative PCR detecting system (qPCR), mass spectrometry, and metagenomic next-generation sequencing (mNGS) have also been applied to detect these fungus. At present, the common clinical identification methods of these fungi are traditional culture methods and Indian ink staining. The clinical fatality rate of these fungi is extremely high and causes more than 180,000 deaths worldwide every year. gattii can easily invade the human central nervous system and cause cryptococcal meningitis (CM). 5Lianyungang Hospital Affiliated to Xuzhou Medical University, Lianyungang, ChinaĬryptococcus neoformans ( C.4Lianyungang Second People’s Hospital Affiliated to Bengbu Medical College, Lianyungang, China.3Department of Materials Science and Engineering, Suzhou University of Science and Technology, Suzhou, China.2School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, China.1Department of Central Laboratory, Lianyungang Hospital Affiliated to Jiangsu University, Lianyungang, China.Lei Wang 1,2†, Yan Wang 1†, Fang Wang 1†, Mengdi Zhao 3, Xuzhu Gao 1, Huimin Chen 1, Na Li 4, Qing Zhu 5, Lipin Liu 5, Wenjun Zhu 1, Xia Liu 1, Yujiao Chen 1, Ping Zhou 1, Yingzhi Lu 1, Kun Wang 1*, Weiguo Zhao 2* and Wei Liang 1*
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