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[采用多重聚合酶链反应法对尸检病例中病毒性呼吸道感染病原体的调查:一项为期五年的研究]

[Investigation of viral respiratory tract infection agents by multiplex PCR method in autopsy cases: A five-year study].

作者信息

Ziyade Nihan, Elgörmüş Neval, Kara Erdoğan, Karayel Ferah

机构信息

Council of Forensic Medicine, Postmortem Microbiology Laboratory, Istanbul, Turkey.

Council of Forensic Medicine, Autopsy Unit, Istanbul, Turkey.

出版信息

Mikrobiyol Bul. 2019 Apr;53(2):179-191. doi: 10.5578/mb.67960.

DOI:10.5578/mb.67960
PMID:31130122
Abstract

Viral respiratory infections are one of the leading causes of morbidity and mortality, especially in children, elderly and immunocompromised patients. The inclusion of post-mortem studies to diagnose the infection causing mortality could be beneficial in specifying new pathogens and determining strategies for treatment and prevention. The aim of this study was to research viral etiology by applying multiplex real-time polymerase chain reaction (Rt-PCR) method in autopsy cases who have been considered to have a respiratory infection and to assess whether the viruses detected are the primary cause of the infection and whether they have any contributory effect on the mortality together with histopathological evidence. In this study, we included a total of 834 cases consisting of sudden death cases from infantile-pediatric age group and autopsy cases from > 18 year age group with suspected respiratory tract infection in our laboratory between January 2013 and May 2017. Of 834 cases, 468 (56.1%) were male and 366 (43.9%) were female, there were 191 (22.9%) cases between 0-1 months, 593 (71.1%) cases between 1 month-18 years, and 50 (6%) cases in the > 18 years age group. In 728 of 834 (87.3%) cases nasopharyngeal/tracheal swab samples and in 106 (12.7%) of them paraffin-embedded lung tissue samples were studied by the use of "FTD Respiratory 21 (Fast-Tract Diagnostics Luxemburg)" kit, with multiplex Rt-PCR method. The post-mortem samples were evaluated for human rhinovirus (HRV), parainfluenza viruses (PIV) (1, 2, 3, 4), influenza virus type A and B (INF-A, INF-B), enterovirus (EV), human bocavirus (HBoV), adenovirus (AdV), human coronavirus (HCoV 229,63,HKU,43), human metapneumovirus A ve B (HMPV-A/B), parechovirus, respiratory syncytial virus (RSV A/B) and Mycoplasma pneumoniae. In our study, at least one respiratory virus was detected by Rt-PCR in 379 (45.4%) of total 834 cases, whereas no viral agent was identified in 455 (54.6%) of the cases. One viral agent was detected in 278 (33.3%), two viral agents were detected in 83 (9.94%) and three viral agents were detected in 18 (2.16%) cases. Overall, the most common viral agent was HRV 110 (13.2%) followed by AdV 39 (4.7%) and RSV A/B 33 (4%). In pediatric cases the rate of positive results for respiratory viruses was 31.8% and in adult group it was 20% (p= 0.032). The most common virus detected among children was HRV and INF-A in adult group. In 101 (12.1%) cases infections caused by two or three agents were diagnosed. Infections with two causative agents were detected as 2.6% (5/191) in 0-1 month age group, 13% (77/593) in 1 month-18 year age group and 2% (1/50) in > 18 age group. The most frequently observed co-infections with double causative agents were HRV and INF-B, HRV and PIV, HRV and HBoV, HRV and AdV combinations. Infections with three causative agents were detected completely among 1 month-18 year age [3% (18/593)] group. In our study, 318 (38.1%) cases had no signs of infection in the postmortem histopathological examination of the lung tissues, while the most common finding was lobular pneumonia/purulent bronchitis in 233 (28%) cases and the second was interstitial pneumonia in 168 (20.1%) cases. When all cases were evaluated in terms of infection, positive results were detected in 469 (56.2%) cases. As a result; postmortem microbiological diagnosis with autopsy and histopathological detection of the patients who are thought to have respiratory tract infection will also determine the infectious agents causing death.

摘要

病毒性呼吸道感染是发病和死亡的主要原因之一,尤其是在儿童、老年人和免疫功能低下的患者中。纳入尸检研究以诊断导致死亡的感染,可能有助于确定新的病原体,并确定治疗和预防策略。本研究的目的是通过多重实时聚合酶链反应(Rt-PCR)方法,对被认为患有呼吸道感染的尸检病例进行病毒病因学研究,并结合组织病理学证据,评估检测到的病毒是否为感染的主要原因,以及它们对死亡率是否有任何促成作用。在本研究中,我们纳入了2013年1月至2017年5月期间在我们实验室的总共834例病例,包括婴儿-儿童年龄组的猝死病例和年龄大于18岁的疑似呼吸道感染的尸检病例。在834例病例中,468例(56.1%)为男性,366例(43.9%)为女性,0-1个月的病例有191例(22.9%),1个月至18岁的病例有593例(71.1%),年龄大于18岁的病例有50例(6%)。在834例病例中的728例(87.3%)中,使用“FTD Respiratory 21(Fast-Tract Diagnostics Luxemburg)”试剂盒,通过多重Rt-PCR方法对鼻咽/气管拭子样本进行了研究;在其中106例(12.7%)中,对石蜡包埋的肺组织样本进行了研究。对尸检样本进行了人类鼻病毒(HRV)、副流感病毒(PIV)(1、2、3、4)、甲型和乙型流感病毒(INF-A、INF-B)、肠道病毒(EV)、人博卡病毒(HBoV)、腺病毒(AdV)、人冠状病毒(HCoV 229、63、HKU、43)、人偏肺病毒A和B(HMPV-A/B)、细小病毒、呼吸道合胞病毒(RSV A/B)和肺炎支原体的检测。在我们的研究中,在834例总病例中的379例(45.4%)中通过Rt-PCR检测到至少一种呼吸道病毒,而在455例(54.6%)病例中未鉴定出病毒病原体。在278例(33.3%)中检测到一种病毒病原体,在83例(9.94%)中检测到两种病毒病原体,在18例(2.16%)中检测到三种病毒病原体。总体而言,最常见的病毒病原体是HRV 110例(13.2%),其次是AdV 39例(4.7%)和RSV A/B 33例(4%)。在儿科病例中,呼吸道病毒检测阳性率为31.8%,在成人组中为20%(p = 0.032)。儿童中检测到的最常见病毒是HRV,成人组中是INF-A。在101例(12.1%)病例中诊断出由两种或三种病原体引起的感染。在0-1个月年龄组中,两种病原体引起的感染检测率为2.6%(5/191),在1个月至18岁年龄组中为13%(77/593),在年龄大于18岁组中为2%(1/50)。最常观察到的两种病原体的合并感染是HRV和INF-B、HRV和PIV、HRV和HBoV、HRV和AdV组合。在1个月至18岁年龄组中完全检测到三种病原体引起的感染[3%(18/593)]。在我们的研究中,318例(38.1%)病例在肺组织的尸检组织病理学检查中没有感染迹象,而最常见的发现是小叶性肺炎/脓性支气管炎233例(28%),其次是间质性肺炎168例(20.1%)。当根据感染对所有病例进行评估时,在469例(56.2%)病例中检测到阳性结果。结果;对被认为患有呼吸道感染的患者进行尸检微生物诊断以及组织病理学检测,也将确定导致死亡的感染病原体。

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