Aksoy Gökmen Ayşegül, Ciçek Candan, Saz Eylem Ulaş, Ozananar Yeliz, Duyu Muhterem
Department of Medical Parasitology, Celal Bayar University Faculty of Medicine, Manisa, Turkey.
Mikrobiyol Bul. 2012 Oct;46(4):614-23.
Human metapneumovirus (hMPV) which is classified in Paramyxoviridae family has been identified in 2001 as the etiological agent of lower respiratory tract infection (LRTI) especially in children. Previous studies indicated that hMPV prevalence in LRTI is between 2-25%, being responsible for 10% of childhood LRTIs and its isolation rate is approximately 6% in hospitalized patients under age three years. The aim of this study was to investigate the hMPV prevalence in children with LRTI in our region. A total of 100 patients (41 female, 59 male) ages between 0-10 years old (median age: 4.8) and who were admitted to Pediatric Clinics of Ege University Medical Faculty Hospital with the diagnosis of LRTI between January-December 2009 were included in the study. Nasopharyngeal swab samples were taken from those patients during the first three days of their symptoms. The presence of hMPV in the samples were investigated by rapid (shell vial) cell culture method using HEp-2 cell line and by real-time reverse transcriptase polymerase chain reaction (rRT-PCR). The methods were performed to the clinical samples simultaneously. In both methods, a standard strain of hMPV provided by Erasmus University was used as positive control and QCMD-2009 hMPV panel was used as external quality control. In our study, 11 and 2 samples were found positive with cell culture and rRT-PCR methods, respectively. Two of rRT-PCR positive samples were also positive in cell culture, while the other nine were positive by only cell culture method. Both of the methods were performed twice due to inconsistent results, however, the same results were obtained in both runs. Studies with QCMD-2009 panel yielded compatible results for five samples, however a positive standard sample (hMPV A subtype, Ct value: 37.31) was found as negative by rRT-PCR test used in this study (RealAccurateTM, Pathofinder, The Netherlands). Our data showed that the prevalence of hMPV detected by rapid cell culture method was 11% in pediatric patients with LRTIs, the age range of hMPV positive cases was 6 months to 7 years old (median age: 20 months), the majority of the admissions was in winter season and the main clinical picture was bronchiolitis. In addition, rRT-PCR assay used in this study was thought to be insufficient to detect the viral RNA in the event of low levels of hMPV A subtypes. Thereby the cell culture method should be used in addition to the new developing molecular methods for the detection of hMPV until standardization is achieved.
人偏肺病毒(hMPV)属于副粘病毒科,于2001年被确认为下呼吸道感染(LRTI)的病原体,尤其是在儿童中。先前的研究表明,hMPV在LRTI中的流行率为2%-25%,占儿童LRTI的10%,在3岁以下住院患者中的分离率约为6%。本研究的目的是调查我们地区LRTI儿童中hMPV的流行情况。共有100例患者(41例女性,59例男性)纳入研究,年龄在0至10岁之间(中位年龄:4.8岁),于2009年1月至12月因LRTI诊断入住伊兹密尔大学医学院附属医院儿科门诊。在症状出现的前三天从这些患者中采集鼻咽拭子样本。使用HEp-2细胞系通过快速(空斑小瓶)细胞培养法和实时逆转录聚合酶链反应(rRT-PCR)检测样本中hMPV的存在。这两种方法同时应用于临床样本。在两种方法中,均使用伊拉斯姆斯大学提供的hMPV标准菌株作为阳性对照,QCMD-2009 hMPV检测板作为外部质量控制。在我们的研究中,细胞培养法和rRT-PCR法分别发现11例和2例样本呈阳性。rRT-PCR阳性的2个样本在细胞培养中也呈阳性,而另外9个样本仅通过细胞培养法呈阳性。由于结果不一致,两种方法均进行了两次,但两次运行均得到相同结果。使用QCMD-2009检测板对5个样本的检测结果相符,但本研究中使用的rRT-PCR检测(RealAccurateTM,Pathofinder,荷兰)将一个阳性标准样本(hMPV A亚型,Ct值:37.31)检测为阴性。我们的数据表明,通过快速细胞培养法检测到的hMPV在LRTI儿科患者中的流行率为11%,hMPV阳性病例的年龄范围为6个月至7岁(中位年龄:20个月),大多数入院发生在冬季,主要临床表现为细支气管炎。此外,本研究中使用的rRT-PCR检测方法在hMPV A亚型水平较低时被认为不足以检测病毒RNA。因此,在实现标准化之前,除了新开发的分子方法外,还应使用细胞培养法来检测hMPV。