Sundaramurthy Raja, Dhodapkar Rahul, Kaliaperumal Subashini, Harish Belgode Narasimha
Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER), Puducherry, India.
J Infect Dev Ctries. 2018 Jan 31;12(1):43-51. doi: 10.3855/jidc.9439.
Highly contagious adenoviral conjunctivitis represents 15-70% of all conjunctivitis worldwide. Human adenovirus (hAdV) serotypes 3,4,7,8,19 and 37 contributes to 89% of all adenoviral conjunctivitis. Accurate and rapid diagnosis of adenoviral infections at serotype level could prevent misdiagnosis, spread of disease, unnecessary antibiotic use and increased treatment costs.
Sixty-two suspected viral conjunctivitis cases were recruited from November2013-January2015. Swabs collected from inferior palpebral conjunctiva and processed for viral culture (Hep2 cell line), immunofluorescence assay (IFA) and polymerase chain reaction (PCR) (targeting hexon gene). Serotype 3,4,7,8,19 and 37 identification was carried out with an optimized multiplex-PCR (based on hypervariable region of hexon gene) and confirmed by sequence analysis. Bayesian Latent Class Model (LCM) analysis was used to compare sensitivity and specificity of three tests.
Adenovirus was detected in 54.8% (34/62) of cases by combination of all three methods. Culture was positive in 23/34 cases (67.6%). PCR and IFA detected adenovirus in 24 (70.5%) and 21 (61.7%) cases respectively. LCM analysis revealed, sensitivity and specificity of PCR, Culture and IFA was 77.8% and 92.4%; 72.2% and 90.8%; 67.6% and 92.9% respectively. Serotyping by multiplex-PCR showed, two cases each were hAdV3 and hAdV4, 18 hAdV8 and two remained unidentified. Results of Multiplex-PCR and sequence analysis showed 100% concordance Conclusion: LCM analysis revealed, PCR is the most appropriate method for identification. Multiplex-PCR is a simple and rapid method (serotypes identification within two days); owing its short turnaround time and accuracy, it can be used as a diagnostic tool for surveillance of adenoviral keratoconjunctivitis.
高传染性腺病毒性结膜炎占全球所有结膜炎病例的15%-70%。人类腺病毒(hAdV)血清型3、4、7、8、19和37导致了89%的腺病毒性结膜炎。在血清型水平上准确快速地诊断腺病毒感染可防止误诊、疾病传播、不必要的抗生素使用以及治疗费用增加。
2013年11月至2015年1月招募了62例疑似病毒性结膜炎病例。从下睑结膜采集拭子并进行病毒培养(Hep2细胞系)、免疫荧光测定(IFA)和聚合酶链反应(PCR)(靶向六邻体基因)。采用优化的多重PCR(基于六邻体基因的高变区)进行血清型3、4、7、8、19和37的鉴定,并通过序列分析进行确认。使用贝叶斯潜在类别模型(LCM)分析来比较三种检测方法的敏感性和特异性。
通过三种方法联合检测,在54.8%(34/62)的病例中检测到腺病毒。培养法在23/34例(67.6%)中呈阳性。PCR和IFA分别在24例(70.5%)和21例(61.7%)中检测到腺病毒。LCM分析显示,PCR、培养法和IFA的敏感性和特异性分别为77.8%和92.4%;72.2%和90.8%;67.6%和92.9%。多重PCR血清分型显示,各有2例为hAdV3和hAdV4,18例为hAdV8,2例仍未鉴定。多重PCR和序列分析结果显示一致性为100%。结论:LCM分析显示,PCR是最合适的鉴定方法。多重PCR是一种简单快速的方法(两天内完成血清型鉴定);由于其周转时间短且准确性高,可作为腺病毒性角结膜炎监测的诊断工具。