Polymerase chain reaction—based assays of vitreous samples for the diagnosis of viral retinitis

Knox, Curtis; Chandler, Diane; Short, Graham; Margolis, Todd P.

doi:10.1016/s0161-6420(98)71127-2

Cited by 143 publications

(82 citation statements)

References 25 publications

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“…PCR analysis of ocular fluid samples may expedite the diagnosis and treatment of such conditions, which is important as the retinitis can progress rapidly. 11,15,16,42,43 Syphilis, once the main cause of vitreous opacities, 1 accounts for only about 1% of all vitreous opacities but is increasing in incidence, particularly in the HIV-positive population, 44 and should always be considered in the differential diagnosis of an inflammatory cellular vitreous infiltrate.…”

Section: Inflammatory Non-infectious Vitritismentioning

confidence: 99%

The pathologist's perspective on vitreous opacities

Coupland

2008

Eye

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Section: Inflammatory Non-infectious Vitritismentioning

confidence: 99%

The pathologist's perspective on vitreous opacities

Coupland

2008

Eye

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“…During the initial stage, only PCR allows rapid and valid results. Time-delayed PCR diagnostics lead to diminished test sensitivity (de Boer et al, 1996;Knox et al, 1998). Due to the high test sensitivity of the PCR, 20-50 μl sample volume is sufficient in most cases.…”

Section: Nucleic Acid Diagnosticsmentioning

confidence: 99%

Virus Diagnostics and Antiviral Therapy in Acute Retinal Necrosis (ARN)

Rautenberg¹,

Hillenkamp²,

Grancicova³

et al. 2012

Antiviral Drugs - Aspects of Clinical Use and Recent Advances

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“…Conventional methods for the diagnosis of viral retinitis include the detection of viral antigen and virus isolation from intraocular specimens (2). These tests have been shown to have low sensitivities for the detection of viruses and are not currently recommended for use for the diagnosis of viral retinitis (2).PCR has proved to be of great utility for the diagnosis of viral retinitis (3,4,5,6,8). However, owing to the expensive systems required, PCR is still not a very common diagnostic test.…”

mentioning

confidence: 99%

“…PCR has proved to be of great utility for the diagnosis of viral retinitis (3,4,5,6,8). However, owing to the expensive systems required, PCR is still not a very common diagnostic test.…”

mentioning

confidence: 99%

Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid and Inexpensive Detection of Cytomegalovirus DNA in Vitreous Specimens from Suspected Cases of Viral Retinitis

et al. 2010

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A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of cytomegalovirus (CMV) was developed and evaluated. The LAMP assay specifically amplified only CMV DNA, and no cross-reactivity with the DNA of herpes simplex virus type 1, varicella-zoster virus, adenovirus, Aspergillus flavus, or Staphylococcus aureus was observed. The sequences of the LAMP assay-positive CMV products were perfectly (100%) matched with the CMV sequence deposited in the GenBank database. The sensitivity of the LAMP assay was found to be 10 copies/l of CMV DNA. Vitreous samples from 40 patients with suspected retinitis were subjected to LAMP and real-time PCR for the detection of CMV. Of 40 patients with suspected viral retinitis, 10 tested positive for CMV by the real-time PCR and LAMP assays. A 100% concordance was observed between the results of the two methods. The LAMP assay is a rapid, highly specific, and sensitive method for the diagnosis of retinitis caused by CMV.Viral retinitis is commonly caused by herpes simplex virus type 1 (HSV-1), HSV-2, varicella-zoster virus (VZV), cytomegalovirus (CMV), and occasionally, Epstein-Barr virus (EBV) (7). In patients with atypical features and in the early stages of ocular manifestations, clinical differentiation between cases of retinitis associated with CMV and other herpesvirus infections is often difficult (6). The differentiation of CMV retinitis from HSV and VZV retinitis is very important early in the course of the disease, as the therapeutic agent to be used for treatment differs from virus to virus (7). Conventional methods for the diagnosis of viral retinitis include the detection of viral antigen and virus isolation from intraocular specimens (2). These tests have been shown to have low sensitivities for the detection of viruses and are not currently recommended for use for the diagnosis of viral retinitis (2).PCR has proved to be of great utility for the diagnosis of viral retinitis (3,4,5,6,8). However, owing to the expensive systems required, PCR is still not a very common diagnostic test. Notomi et al. have reported on a novel nucleic acid amplification assay termed the loop-mediated isothermal amplification (LAMP) assay (10). The assay amplifies the DNA under isothermal conditions (63 to 65°C) with high degrees of specificity, efficiency, and speed. The assay can be conducted in the laboratory in a water bath or heating block (10). Thus, the thermal cycling needs of a PCR are avoided. The assay can be used for the rapid detection of pathogens in peripheral health care settings in developing countries. The present study describes the development and evaluation of a simple and costeffective LAMP assay for the rapid detection of CMV DNA in patients with viral retinitis. MATERIALS AND METHODSClinical samples, positive controls, and DNA extraction. The study was approved (reference no. LEC 08013) by the Institutional Review Board of the L. V. Prasad Eye Institute, Hyderabad, India. Vitreous samples from 40 patients with viral retinitis were included in the st...

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Polymerase chain reaction—based assays of vitreous samples for the diagnosis of viral retinitis

Cited by 143 publications

References 25 publications

The pathologist's perspective on vitreous opacities

The pathologist's perspective on vitreous opacities

Virus Diagnostics and Antiviral Therapy in Acute Retinal Necrosis (ARN)

Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid and Inexpensive Detection of Cytomegalovirus DNA in Vitreous Specimens from Suspected Cases of Viral Retinitis

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