Marova A A, Oksanich A S, Kaira A N, Meskina E R, Medvedeva E A, Ivanova O E, Lukashev A N, Kyuregian K K, Kalinkina M A, Egorova O V, Zverev V V, Faĭzuloev E V
Zh Mikrobiol Epidemiol Immunobiol. 2012 Nov-Dec(6):39-45.
Evaluate the effectiveness of multiplex reverse transcription (RT) and polymerase chain reaction with fluorescence detection in real time mode (qPCR) methods for differential detection of 11 groups of intestine viruses (adenoviruses, enteroviruses, polioviruses, hepatitis A and E viruses, group A and C rotaviruses, orthoreoviruses, noroviruses, sapoviruses and astroviruses) in various biological samples.
Panels of virus isolates and clinical samples characterized by reference methods were used to evaluate sensitivity of detection of various intestine viruses. Nucleic acids were isolated from study samples and multiplex RT and qPCR were carried out.
Sensitivity of laboratory reagent kit (LRK) when compared with results obtained from reference methods was 100% for rotavirus A, adenovirus, enterovirus and norovirus, 88.9% for hepatitis E virus and 92.3% for hepatitis A virus, and diagnostic specificity - 99.4%. During analysis of 697 clinical samples from patients with acute intestine infection symptoms nucleic acids of various intestine viruses were isolated in 71.7%.
Multiplex qRT-PCR was shown as an effective method of etiologic diagnostics of an intestine viral infection. Use of LRK was demonstrated to establish etiology of intestine diseases in 63 - 72% and in children with watery diarrhea - in approximately 90% of cases.
评估多重逆转录（RT）和实时荧光检测聚合酶链反应（qPCR）方法在不同生物样本中对11组肠道病毒（腺病毒、肠道病毒、脊髓灰质炎病毒、甲型和戊型肝炎病毒、A组和C组轮状病毒、正呼肠孤病毒、诺如病毒、札如病毒和星状病毒）进行鉴别检测的有效性。
使用经参考方法鉴定的病毒分离株和临床样本评估不同肠道病毒检测的敏感性。从研究样本中分离核酸并进行多重RT和qPCR。
与参考方法获得的结果相比，实验室试剂试剂盒（LRK）对A组轮状病毒、腺病毒、肠道病毒和诺如病毒的敏感性为100%，对戊型肝炎病毒为88.9%，对甲型肝炎病毒为92.3%，诊断特异性为99.4%。在对697例有急性肠道感染症状患者的临床样本进行分析时，71.7%的样本中分离出了各种肠道病毒的核酸。
多重qRT-PCR被证明是肠道病毒感染病因诊断的有效方法。使用LRK可在63 - 72%的病例中确定肠道疾病的病因，在儿童水样腹泻病例中约90%的病例可确定病因。
