Chichili Gurunadh Reddy, Athmanathan Sreedharan, Farhatullah Syed, Gangopadhyay Nibaran, Jalali Subhadra, Pasricha Gunisha, Sharma Savitri
Jhaveri Microbiology Center, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, L.V Prasad Marg, Banjara Hills, Hyderabad, India.
Curr Eye Res. 2003 Aug;27(2):85-90. doi: 10.1076/ceyr.27.2.85.15947.
A majority of ocular viral diseases are caused by herpes group of viruses. Such infections, especially atypical herpetic keratitis, iridocyclitis and intra-ocular inflammations, can often present with overlapping clinical manifestations misleading the diagnosis. Molecular techniques are most useful in such instances for an accurate and rapid diagnosis since conventional methods are time consuming and less sensitive. A multiplex PCR was developed and used for the detection of herpes simplex virus (HSV), varicella zoster virus (VZV), and cytomegalovirus (CMV) in ocular samples.
One hundred and forty six ocular samples (corneal scrapings - 52, aqueous fluid - 36, vitreous fluid - 31, tissues - 26, skin vesicle scraping - 1) were included in the study. The sensitivity of the assay was determined using serial dilutions of standard strains of HSV, VZV, and CMV vis-à-vis plaque forming assay.
The sensitivity of the assay was 4, 4 and 12 PFU/ml or 20, 20 and 60 genome copy numbers of HSV, VZV and CMV respectively. Using DNA from various sources (fungal, bacterial, human leukocytes, tissues) along with standard positive controls, the assay was found to be highly specific. HSV DNA was detected in majority of the clinical samples (33.6%), most frequent being corneal samples. Comparatively, VZV and CMV infections were detected in small number of samples (VZV-3, CMV-2).
We found the assay very useful in our set-up whenever a differential diagnosis of herpetic infections was suggested by the ophthalmologist. The multiplex PCR we have described here can be of greater value in clinics with larger number of patients suspected of having HSV, VZV or CMV infections.
大多数眼部病毒性疾病由疱疹病毒组引起。此类感染，尤其是非典型疱疹性角膜炎、虹膜睫状体炎和眼内炎症，常常表现出重叠的临床表现，从而误导诊断。由于传统方法耗时且敏感性较低，分子技术在此类情况下对于准确快速的诊断最为有用。开发了一种多重聚合酶链反应（PCR）并用于检测眼部样本中的单纯疱疹病毒（HSV）、水痘带状疱疹病毒（VZV）和巨细胞病毒（CMV）。
本研究纳入了146份眼部样本（角膜刮片 - 52份、房水 - 36份、玻璃体 - 31份、组织 - 26份、皮肤水疱刮片 - 1份）。通过对HSV、VZV和CMV标准菌株进行系列稀释并与空斑形成试验相对照，确定了该检测方法的敏感性。
该检测方法的敏感性分别为HSV 4 PFU/ml、VZV 4 PFU/ml和CMV 12 PFU/ml，或分别为HSV、VZV和CMV的20、20和60个基因组拷贝数。使用来自各种来源（真菌、细菌、人白细胞、组织）的DNA以及标准阳性对照，发现该检测方法具有高度特异性。在大多数临床样本（33.6%）中检测到HSV DNA，最常见于角膜样本。相比之下，在少数样本中检测到VZV和CMV感染（VZV - 3例，CMV - 2例）。
我们发现，每当眼科医生提出疱疹感染的鉴别诊断时，该检测方法在我们的机构中非常有用。我们在此描述的多重PCR在怀疑患有HSV、VZV或CMV感染的患者数量较多的诊所中可能具有更大的价值。
