脓毒症患者血疱疹病毒核酸检测阳性的高危因素和临床特征

Risk factors and clinical features of septic patients with human herpes viruses' nucleic acid detected positive in blood

目的利用微生物组学第二代基因测序(next generation sequencing, NGS)技术,测定脓毒症患者外周血疱疹病毒(human herpes Virus,HHV)的检出序列数,探讨HHV检出序列数与脓毒症患者病情程度、预后以及免疫功能状态的相关性。方法收集150例脓毒症患者的血标本和临床信息,进行病原体微生物二代测序,根据是否检出HHV核酸将患者分为检出组和未检出组。在采集血标本当天进行序贯器官衰竭评估(sequential organ failure assessment, SOFA)和急性生理慢性健康评分Ⅱ(acute physiology and chronic health evaluation Ⅱ, APACHE Ⅱ),并测定外周血白细胞和淋巴细胞计数、淋巴细胞分类,用固相夹心、化学发光法测定血清中细胞因子的浓度。结果 51.3%的脓毒症患者血中检出HHV核酸;HHV检出组患者APACHE Ⅱ和SOFA评分明显高于未检出组,28 d和90 d病死率也明显高于未检出组,差异有统计学意义(P<0.01)。脓毒症患者病情严重程度与HHV检出率成正相关。脓毒症患者HHV核酸检出组与未检出组相比,住院90 d病死率呈正相关(P=0.0056)。单因素分析发现检出组患者外周血淋巴细胞计数及CD19+B、CD4+T、CD8+T、CD56+淋巴细胞显著低于未检出组;检出组中促炎因子(TNF-α、IL-2R、IL-6、IL-8)和抗炎因子(IL-10)水平均高于未检出组(P<0.01)。Logistic多因素回归分析去除混杂因素后,性别、年龄、APACHE Ⅱ和SOFA评分、IL-2R、IL-10、CD19+B淋巴细胞、T淋巴细胞计数仍然与HHV检出阳性有关。结论脓毒症患者人疱疹病毒DNA的检出率较高,外周血检出HHV核酸序列是患者预后不良的高危因素;将HHV核酸序列数检出值定为100/样本判断患者的预后,有显著的临床价值。HHV检出与患者的免疫功能状态有关。

Objective To measure the reads numbers of Human Herpes Virus in blood sample from patients with sepsis by using Next Generation sequencing (NGS) and explore the relationship between read number of virus and the severity, prognosis, immune status of septic patients. Methods Blood sample and clinical information from 150 patients with sepsis were enrolled in this study. All patients' blood samples were sent to perform NGS pathogenic test. According to the results of NGS, septic patients were divided into HHV-detected group and HHV-undetected group. Besides, patients were scored with Sequential Organ Failure Assessment (SOFA) and Acute Physiology and Chronic Health Evaluation Ⅱ(APACHE Ⅱ) on the day of blood collection. The counts of total leukocytes, lymphocytes and the levels of cytokines were also measured. Results 51.3 percent of septic patients were detected with HHV nucleic acid. The APACHE Ⅱ and SOFA scores were significantly higher in HHV-detected patients compared with patients in HHV-undetected group. Besides, patients who had a higher SOFA score might lead to a higher detection rate of HHV. Moreover, the 28-day and 90-day mortality rates were higher in detected group (P < 0.01). The detection of HHV nucleic acid was positively correlated with a high 90-day mortality rate (P=0.0056). One-way analysis of variance revealed that the counts of total lymphocyte and different types of lymphocyte (CD19+B、CD4+T、CD8+T、CD56+ lymphocyte) were significantly less in detected group than that in undetected group. Furthermore, both the levels of pro-inflammatory cytokines (TNF-α、IL-2R、IL-6、IL-8) and anti-inflammatory cytokines (IL-10) in detected group were significantly higher than those in undetected group. Gender, age, APACHEⅡ, SOFA, IL-2R, IL-10, CD19+B lymphocyte and T cells, were still significant even after multivariate logistic analyses. Conclusions The detection rate of HHV nucleic acid in patients with sepsis was high. The detection of HHV was a high-risk factor of death in patients with sepsis. The cut-off value which is more than 100 had a significant clinical value. The infection of HHV could be conducted by dysfunction of immunity.