Sultani Mozhdeh, Mokhtari Azad Talat, Eshragian Mohammadreza, Shadab Azadeh, Naseri Maryam, Eilami Owrang, Yavarian Jila
Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran.
Jundishapur J Microbiol. 2015 Aug 1;8(8):e19041. doi: 10.5812/jjm.19041v2. eCollection 2015 Aug.
It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently.
The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR).
Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43.
Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012).
Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.
医生常常难以区分呼吸道感染的病毒病因和细菌病因，这可能导致抗生素的过度使用。在许多社区获得性呼吸道感染病例中，临床医生会进行经验性治疗。近年来，直接检测病毒的分子方法有了进展。
本研究的目的是通过多重SYBR Green实时聚合酶链反应（PCR）识别呼吸道RNA病毒组。
收集了172份住院患者各年龄组的流感阴性呼吸道标本。提取RNA后，合成cDNA。开发了三种SYBR Green多重实时PCR检测方法，用于同时检测12种呼吸道RNA病毒。每组多重方法检测四种病毒，第一组：呼吸道合胞病毒、人偏肺病毒、鼻病毒、肠道病毒；第二组：副流感病毒1 - 4型（PIV1 - 4）；第三组：冠状病毒NL63、229E、严重急性呼吸综合征（SARS）和OC43。
在为期一年的研究中，对172例患者的临床样本应用多重SYBR Green实时PCR检测，结果检测到19例（11.04%）PIV3、9例（5.23%）PIV4和1例（0.58%）冠状病毒NL63。所有阳性样本均在（2011 - 2012年）12月至3月期间检测到。
多重SYBR Green实时PCR是一种快速且相对廉价的呼吸道病毒检测方法。
