Robert Koch-Institut, Nationales Referenzzentrum für Influenza, Nordufer 20, 13353 Berlin, Germany.
J Clin Virol. 2010 Apr;47(4):366-71. doi: 10.1016/j.jcv.2010.01.005. Epub 2010 Feb 10.
In respiratory infections, human adenoviruses (hAdV) of species B1 and C are frequently detected, but severe or even fatal disease outbreaks are predominantly caused by only few serotypes. The molecular typing of hAdV hexon sequences can help to speed up the discrimination of serotypes, thus improving on-time epidemiological examinations and patient care.
We aimed to develop a molecular method for the rapid species B1 and C serotype identification in respiratory samples based on sequence generation of the hexon hypervariable region (HVR).
We developed two PCR-based genotyping systems for a generic HVR amplification and sequence determination of species B1 and C viruses. The assays were applied to hAdV prototypes and 106 samples.
The primer sets proved to be capable of amplifying all B1 and C serotypes. The viruses detected in clinical samples belong to serotypes 1, 2, 3, 5 and 6. The obtained sequences of serotypes 2, 3 and 5 form 2-3 phylogenetic clusters that are based on the characteristic amino acid changes within the variable HVR sites.
Our assay can significantly speed up the time-span needed for serotype identification and will improve epidemiological surveillance and patient care. The obtained hexon sequences of field viruses vary significantly and form multiple genetic lineages. The variability is focussed on the HVR sites and can be interpreted as the ongoing evolutionary process. Further research is needed on the hexon sequence variability of other (respiratory) hAdV serotypes.
在呼吸道感染中,常可检测到物种 B1 和 C 的人腺病毒(hAdV),但严重甚至致命的疾病暴发主要由少数几个血清型引起。hAdV 六邻体序列的分子分型有助于加快血清型的区分,从而改善及时的流行病学检查和患者护理。
我们旨在开发一种基于六邻体超变区(HVR)序列生成的快速鉴定呼吸道样本中物种 B1 和 C 血清型的分子方法。
我们开发了两种基于 PCR 的基因分型系统,用于通用 HVR 扩增和 B1 和 C 病毒的序列测定。该检测方法应用于 hAdV 原型和 106 个样本。
引物组被证明能够扩增所有 B1 和 C 血清型。从临床样本中检测到的病毒属于血清型 1、2、3、5 和 6。获得的血清型 2、3 和 5 的序列形成 2-3 个基于可变 HVR 位点特征氨基酸变化的系统发育簇。
我们的检测方法可以显著缩短血清型鉴定所需的时间,并将改善流行病学监测和患者护理。从现场病毒获得的六邻体序列变化很大,并形成多个遗传谱系。变异性集中在 HVR 位点,可以解释为正在进行的进化过程。需要对其他(呼吸道)hAdV 血清型的六邻体序列变异性进行进一步研究。