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[下呼吸道感染儿童腺病毒的调查]

[Investigation of adenoviruses in children with lower respiratory tract infections].

作者信息

Sağlık Imran, Mutlu Derya, Ongüt Gözde, Velipaşaoğlu Güney Sevtap, Ozkul Aykut, Oğünç Dilara, Colak Dilek

机构信息

Department of Medical Microbiology, Akdeniz University Faculty of Medicine, Antalya, Turkey.

出版信息

Mikrobiyol Bul. 2013 Apr;47(2):282-94. doi: 10.5578/mb.4773.

Abstract

Adenoviruses which are one of the causative agents of acute respiratory tract infections at all age groups worldwide, can lead to epidemic, endemic or sporadic infections year-round. Adenovirus infections in lower respiratory tract can be presented as bronchitis, bronchiolitis and pneumonia. The aim of this study was to investigate the presence of adenoviruses as the etiologic agent of lower respiratory tract infections (LRTIs) in children by cell culture, polymerase chain reaction (PCR) and direct fluorescence antibody (DFA) test. The results of the laboratory tests were evaluated in the light of patients' clinical findings. The study consisted of 206 patients aged between 0-5 years old and who were admitted to the hospital with the complaints of LRTI between January 2011 and April 2012. The clinical, radiological and laboratory findings of the patients were recorded. Nasopharyngeal specimens were taken with flocked swab from all patients and adenoviruses were investigated by shell-vial cell culture, real-time PCR and DFA test, simultaneously. Of all the samples 89.3% were taken in January, February and March and 38% of the patients have one or more chronic underlying diseases as chromosomal abnormalities, congenital heart disease, heart failure, asthma, cystic fibrosis, leukemia, kidney failure and prematurity. Adenovirus was detected in 12 (5.8%) of the samples by PCR, seven (3.4%) of the samples by cell culture method. While seven samples were found positive with both PCR and cell culture, 194 samples yielded negative results in both tests. Five samples, which were found positive by PCR, were not grown in cell culture method. Twelve of the 153 samples examined with DFA test, could not be evaluated due to insufficient amount of cells, however 2.8% (4/141) of the samples were found positive for adenovirus antigens by DFA method. Those samples were also positive ones in the other two methods. Compared with cell culture, the sensitivity, specificity, positive and negative predictive values of PCR were 100%, 97.5%, 58.3% and 100%, respectively; those values were 57%,100%,100% and 97.7%, respectively for DFA testing. Compared to PCR the sensitivity of cell culture is very low (16.6%) after three days of incubation, however, it increased to 58.3% after five days' of incubation. There was no significant relationship between adenovirus positivity and the presence of chronic diseases, the radiological findings and the laboratory findings. Of all adenovirus positive samples 83.3% were obtained in January, February and March. Our data indicated that the etiological agent was adenovirus in approximately 6% of children with LRTI. The most important step for the isolation of adenovirus from respiratory tract, is high quality and sufficient amounts of sample. The flexible flocked swabs made it easy to take nasopharyngeal swab from children. Although cell culture is still the gold standard for the diagnosis of adenovirus infections, PCR which is a fast method with high sensitivity and specificity can also be used. However, specific care should be taken during the DNA extraction stage, since the amount of the nucleic acid in the sample is critical for the best results. Even though the low sensitivity of DFA restricts its use in routine diagnosis of adenovirus infections, it should always be kept in mind that the quality of the clinical samples is most reliably evaluated by this method.

摘要

腺病毒是全球所有年龄组急性呼吸道感染的病原体之一,可全年导致流行、地方性或散发性感染。下呼吸道腺病毒感染可表现为支气管炎、细支气管炎和肺炎。本研究的目的是通过细胞培养、聚合酶链反应(PCR)和直接荧光抗体(DFA)试验,调查腺病毒作为儿童下呼吸道感染(LRTIs)病原体的存在情况。根据患者的临床发现对实验室检查结果进行评估。该研究包括206例年龄在0至5岁之间的患者,他们于2011年1月至2012年4月因LRTI症状入院。记录了患者的临床、放射学和实验室检查结果。用植绒拭子从所有患者采集鼻咽标本,并同时通过空斑小室细胞培养、实时PCR和DFA试验检测腺病毒。所有样本中89.3%在1月、2月和3月采集,38%的患者有一种或多种慢性基础疾病,如染色体异常、先天性心脏病、心力衰竭、哮喘、囊性纤维化、白血病、肾衰竭和早产。通过PCR在12份(5.8%)样本中检测到腺病毒,通过细胞培养法在7份(3.4%)样本中检测到腺病毒。虽然7份样本PCR和细胞培养均呈阳性,但194份样本两种检测均为阴性。5份PCR检测呈阳性的样本在细胞培养法中未生长。用DFA试验检测的153份样本中有12份因细胞数量不足无法评估,然而,通过DFA法在2.8%(4/141)的样本中发现腺病毒抗原呈阳性。这些样本在其他两种方法中也呈阳性。与细胞培养相比,PCR的敏感性、特异性、阳性和阴性预测值分别为100%、97.5%、58.3%和100%;DFA检测的这些值分别为57%、100%、100%和97.7%。与PCR相比,细胞培养在孵育三天后的敏感性非常低(16.6%),然而,在孵育五天后增加到58.3%。腺病毒阳性与慢性病的存在、放射学检查结果和实验室检查结果之间无显著关系。所有腺病毒阳性样本中83.3%在1月、2月和3月获得。我们的数据表明,在约6%的LRTI儿童中病原体为腺病毒。从呼吸道分离腺病毒的最重要步骤是高质量和足够数量的样本。柔性植绒拭子便于从儿童采集鼻咽拭子。虽然细胞培养仍然是腺病毒感染诊断的金标准,但PCR作为一种快速、高灵敏度和特异性的方法也可使用。然而,在DNA提取阶段应特别注意,因为样本中的核酸量对获得最佳结果至关重要。尽管DFA的低灵敏度限制了其在腺病毒感染常规诊断中的应用,但应始终牢记,通过该方法可最可靠地评估临床样本的质量。

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