四种不同检测方法在人类免疫缺陷病毒感染合并隐球菌脑膜炎中的诊断价值

Diagnostic value of 4 different detection methods in human immunodeficiency virus infection complicated with cryptococcal meningitis

目的探讨在脑脊液(cerebro-spinal fluid,CSF)样本中,隐球菌荚膜抗原胶体金免疫层析法、墨汁染色法、真菌培养法以及病原宏基因组二代测序(metagenomic next-generation sequencing,mNGS)技术诊断HIV感染合并隐球菌脑膜炎(HIV-cryptococcal meningitis,HIV-CM)疾病的价值。方法收集并分析2020年12月—2022年12月武汉大学中南医院感染科收治的171例考虑中枢神经系统感染行腰椎穿刺进行隐球菌检测的HIV/AIDS住院患者的一般及临床资料,并比较4种检测方法对HIV-CM的检测阳性率、敏感度和特异度。结果171例研究对象中最终诊断为HIV-CM的患者共35例。HIV-CM患者以神经系统受累,伴发热、头晕、头痛为主要表现。171例脑脊液样本中,隐球菌荚膜抗原胶体金免疫层析法检测阳性率为18.9%,墨汁染色法检测阳性率为15.4%,真菌培养法检测阳性率为10.7%,mNGS技术检测阳性率为23.3%。隐球菌荚膜抗原胶体金免疫层析法、墨汁染色法、真菌培养法、mNGS技术检测敏感度分别为97.0%、75.8%、52.9%和100%,特异度分别为100%、99.3%、100%和100%。4种检测方法脑脊液样本送检至报告的平均时间分别为0.5 d、0.5~1 d、5 d和2 d。10例HIV-CM患者mNGS检测结果显示同时有3种及以上病原菌多重感染的患者有7例。结论脑脊液样本的墨汁染色检测可为早期筛查获得病原学诊断依据,真菌培养法是诊断金标准且培养菌落可用于后续药物敏感性试验,隐球菌抗原胶体金免疫层析法和mNGS技术的检测敏感度和特异度都高,报告周期短,且mNGS技术可用于HIV合并多重感染患者的早期病原学诊断。因此,建议尽量联合多方法同时检测,为HIV-CM的临床诊断提供更准确更快速的临床证据,为患者争取更大获益。

Objective To explore the diagnostic value of cryptococcal antigen,India ink staining,fungal culture,and metagenomic next-generation sequencing(mNGS)of in cerebrospinal fluid(CSF)samples in HIV associated cryptococcal meningitis(HIV-CM).Method A retrospective analysis was conducted on the clinical data of 171 HIV/AIDS hospitalized patients with central nervous system infection who underwent lumbar puncture for cryptococcal testing in the Department of Infectious Diseases,Zhongnan Hospital of Wuhan University,from December 2020 to December 2022.General information,clinical manifestations,and laboratory test results of HIV-CM patients were collected,and the positive rate,sensitivity,and specificity of the four detection methods for HIVCM were compared.Results Among the 171 study subjects,a total of 35 patients were ultimately diagnosed with HIV-CM.The main manifestations of HIV-CM patients were neurological involvement,accompanied by fever,dizziness,and headache.Among 171 cerebrospinal fluid samples,the positive rate of CrAg detection was 18.9%,ink staining was 15.4%,fungal culture was 10.7%,and mNGS was 23.3%.The sensitivity of the CrAg detection,ink staining,fungal culture,and mNGS were 97.0%,75.8%,52.9%and 100%,respectively,and the specificity were 100%,99.3%,100%and 100%,respectively.The average reported time is 0.5 d,0.5-1 d,5 d and 2 d,respectively.The mNGS test results of 10 patients showed that up to 7 patients had multiple infections of three or more pathogenic bacteria simultaneously.Conclusions Ink staining of CSF samples can provide early screening for pathogenic diagnosis.Fungal culture is the gold standard for diagnosis,and subsequent drug sensitivity testing is needed.Cryptococcus antigen detection and mNGS have high sensitivity and specificity,and the detection report time is short.mNGS can be used for early pathogenic diagnosis of HIV patients with multiple infections.Therefore,it is recommended to combine various methods for simultaneous testing as much as possible to provide more accurate and rapid clinical evidence for the diagnosis of HIV-CM and to strive for greater benefits for patients.