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血清学检测指标及实时荧光定量PCR检测在侵袭性念珠菌病患者中的诊断价值

Diagnostic Value of Serological Markers and Quantitative Real-Time PCR in Patients with Invasive Candidiasis

    摘要
  • 摘要:
    目的 评估血清学检测指标1, 3-β-D-葡聚糖试验（G试验）、念珠菌甘露聚糖（Mn）抗原、念珠菌甘露聚糖IgG抗体（Mn-IgG）和实时荧光定量PCR（qPCR）检测在侵袭性念珠菌病（IC）患者中的临床诊断价值。
    方法 收集四川大学华西医院2022年12月-2024年12月期间通过血培养及无菌体液培养确诊为IC患者的血清样本79例作为IC组，排除IC感染患者的血清样本83例作为非IC组，进行G试验（磁微粒化学发光法）、Mn抗原（ELISA法）、Mn-IgG抗体（化学发光法）和qPCR检测。比较分析G试验、Mn抗原、Mn-IgG抗体和qPCR检测及联合检测对IC诊断的敏感性、特异性、阳性预测值、阴性预测值，绘制受试者工作特征（ROC）曲线并计算ROC曲线下面积（AUC）。
    结果 3种血清学检测指标单独检测对IC诊断的敏感性、特异性、阳性预测值、阴性预测值分别为：G试验62.03%、94.87%、92.45%、71.15%；Mn抗原65.63%、86.59%、79.25%、76.34%；Mn-IgG抗体31.65%、87.95%、71.43%、57.48%。G试验、Mn抗原、Mn-IgG三项联合检测敏感性提高至73.44%。基于ROC曲线分析，各血清学检测指标诊断IC的AUC分别为：G试验 0.862 (95% CI: 0.803-0.922)、Mn试验 0.853 (95% CI: 0.790-0.915)、Mn-IgG 0.603 (95% CI: 0.514-0.692)。G试验+Mn试验两项联合检测、G试验+Mn-IgG两项联合检测、G试验+Mn试验+Mn-IgG三项联合检测的AUC分别为0.875（95% CI: 0.809-0.925）、0.869（95% CI: 0.805-0.917）和0.875（95% CI: 0.809-0.924），优于单独检测。qPCR对IC诊断的敏感性、特异性、阳性预测值、阴性预测值分别为100.00%、98.80%、98.75%、100.00%。以培养菌株基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF MS）鉴定结果为金标准，qPCR检测对白念珠菌、热带念珠菌、光滑念珠菌、近平滑念珠菌的诊断一致性分别为100.00%、100.00%、94.44%和90.00%。
    结论 G试验、Mn抗原和Mn-IgG抗体联合检测可提高诊断敏感性，有助于IC的早期诊断。qPCR对IC具有优异的诊断性能，可将IC常见念珠菌鉴定至种水平，并可覆盖其他念珠菌种，与临床培养方法结果一致性好。

     

    Abstract:
    Objective  To evaluate the clinical diagnostic value of serological markers, including the 1, 3-β-D-glucan test (G-test), Candida mannan (Mn) antigen, Candida mannan IgG antibody (Mn-IgG), and quantitative real-time PCR(qPCR) in patients with invasive candidiasis (IC).
    Methods  This study included 79 patients with culture-proven IC (IC group) and 83 patients without IC (non-IC group) at West China Hospital, Sichuan University between December 2022 and December 2024. Serum samples were tested for the G test (chemiluminescence), Mn antigen (ELISA), Mn-IgG (chemiluminescence), and qPCR. The diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of individual and combined tests were calculated. Diagnostic performance was assessed using receiver operating characteristic (ROC) curve analysis.
    Results The sensitivity, specificity, PPV, and NPV of the three serological markers for IC diagnosis when used individually were as follows: G test, 62.03%, 94.87%, 92.45%, and 71.15%; Mn antigen, 65.63%, 86.59%, 79.25%, and 76.34%; Mn-IgG antibody, 31.65%, 87.95%, 71.43%, and 57.48%. Combined detection of the G test, Mn antigen, and Mn-IgG increased sensitivity to 73.44%. Based on ROC curve analysis, the AUCs for diagnosing IC using each serological marker were: G test, 0.862 (95% CI: 0.803-0.922); Mn test, 0.853 (95% CI: 0.790-0.915); Mn-IgG, 0.603 (95% CI: 0.514-0.692). The combined AUCs for G test + Mn test, G test + Mn-IgG, and G test + Mn test + Mn-IgG were 0.875 (95% CI: 0.809-0.925), 0.869 (95% CI: 0.805-0.917), and 0.875 (95% CI: 0.809-0.924), respectively, demonstrating superiority over individual tests. The sensitivity, specificity, PPV, and NPV of qPCR for IC diagnosis were 100.00%, 98.80%, 98.75%, and 100.00%, respectively. Using culture strain identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as the gold standard, qPCR demonstrated diagnostic concordance rates of 100.00% for Candida albicans, 100.00% for Candida tropicalis, 94.40% for Candida glabrata, and 90.00% for Candida parapsilosis.
    Conclusion The combined use of the G test, Mn antigen, and Mn-IgG antibody increases diagnostic sensitivity and facilitates early diagnosis of IC. qPCR demonstrates excellent diagnostic performance for IC, enabling species-level identification of common Candida species associated with IC and detection of other Candida species. It also shows consistency with clinical culture methods.

     

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