非 HIV 患者中肺孢子菌肺炎的临床特征与液滴数字 PCR 拷贝数的关系。
Relationship between clinical features and droplet digital PCR copy number in non-HIV patients with pneumocystis pneumonia.
机构信息
Department of Respiratory and Critical Care Medicine, Peking University People's Hospital, No.11, Xizhimen South Street, Beijing, China.
Department of Respiratory Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
出版信息
BMC Infect Dis. 2023 Nov 27;23(1):833. doi: 10.1186/s12879-023-08580-7.
OBJECTIVE
Droplet digital PCR (ddPCR) is a novel assay to detect pneumocystis jjrovecii (Pj) which has been defined to be more sensitive than qPCR in recent studies. We aimed to explore whether clinical features of pneumocystis pneumonia (PCP) were associated with ddPCR copy numbers of Pj.
METHODS
A total of 48 PCP patients were retrospectively included. Pj detection was implemented by ddPCR assay within 4 h. Bronchoalveolar fluid (BALF) samples were collected from 48 patients with molecular diagnosis as PCP via metagenomic next generation sequencing (mNGS) or quantitative PCR detection. Univariate and multivariate logistic regression were performed to screen out possible indicators for the severity of PCP. The patients were divided into two groups according to ddPCR copy numbers, and their clinical features were further analyzed.
RESULTS
Pj loading was a pro rata increase with serum (1,3)-beta-D glucan, D-dimmer, neutrophil percentage, procalcitonin and BALF polymorphonuclear leucocyte percentage, while negative correlation with albumin, PaO2/FiO2, BALF cell count, and BALF lymphocyte percentage. D-dimmer and ddPCR copy number of Pj were independent indicators for moderate/severe PCP patients with PaO2/FiO2 lower than 300. We made a ROC analysis of ddPCR copy number of Pj for PaO2/FiO2 index and grouped the patients according to the cut-off value (2.75). The high copy numbers group was characterized by higher level of inflammatory markers. Compared to low copy number group, there was lower level of the total cell count while higher level of polymorphonuclear leucocyte percentage in BALF in the high copy numbers group. Different from patients with high copy numbers, those with high copy numbers had a tendency to develop more severe complications and required advanced respiratory support.
CONCLUSION
The scenarios of patients infected with high ddPCR copy numbers of Pj showed more adverse clinical conditions. Pj loading could reflect the severity of PCP to some extent.
目的
液滴数字 PCR（ddPCR）是一种新型检测方法，用于检测卡氏肺孢子虫（Pj），最近的研究表明，ddPCR 比 qPCR 更灵敏。我们旨在探讨卡氏肺孢子虫肺炎（PCP）的临床特征是否与 Pj 的 ddPCR 拷贝数有关。
方法
回顾性纳入 48 例 PCP 患者。通过 ddPCR 检测在 4 小时内进行 Pj 检测。通过宏基因组下一代测序（mNGS）或定量 PCR 检测对 48 例分子诊断为 PCP 的患者采集支气管肺泡灌洗液（BALF）样本。采用单因素和多因素 logistic 回归筛选可能与 PCP 严重程度相关的指标。根据 ddPCR 拷贝数将患者分为两组，并进一步分析其临床特征。
结果
Pj 负荷与血清（1,3）-β-D 葡聚糖、D-二聚体、中性粒细胞百分比、降钙素原和 BALF 中性粒细胞百分比呈比例增加，与白蛋白、PaO2/FiO2、BALF 细胞计数和 BALF 淋巴细胞百分比呈负相关。D-二聚体和 Pj 的 ddPCR 拷贝数是 PaO2/FiO2 低于 300 的中重度 PCP 患者的独立指标。我们对 Pj 的 ddPCR 拷贝数进行了 PaO2/FiO2 指数的 ROC 分析，并根据截断值（2.75）将患者分组。高拷贝数组的炎症标志物水平更高。与低拷贝数组相比，高拷贝数组 BALF 中的总细胞计数较低，而中性粒细胞百分比较高。与高拷贝数组患者不同，高拷贝数组患者有发展为更严重并发症的趋势，需要更先进的呼吸支持。
结论
Pj 的 ddPCR 拷贝数较高的患者病情更差。Pj 负荷在一定程度上可以反映 PCP 的严重程度。
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