Detection of West Nile virus using formalin fixed paraffin embedded tissues in crows and horses: quantification of viral transcripts by real-time RT-PCR

Tewari, Deepanker; Kim, Hyun; Feria, Willard; Russo, Brigite; Acland, Helen M.

doi:10.1016/j.jcv.2004.01.003

Cited by 14 publications

(6 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although studies showed that brain tissue[4] and intestine[5] have been the most reliable organ for VI and RT-PCR testing of crows for WNv, this study showed that the spleen appears to be the most reliable organ for IHC testing. Since in terms of practicality, multiple organs can be screened with the same expense and time, it is recommended that snippets of liver, spleen, kidney, lung, and intestine be included to increase the accuracy of result.…”

Section: Discussioncontrasting

confidence: 57%

Antigenic distribution of west nile virus in various organs of wildly infected American crows (Corvus Brachyrhynchos)

Sandhu¹,

Sidhu

Dhillon³

2011

J Global Infect Dis

View full text Add to dashboard Cite

Objective:Since its discovery in the western hemisphere in 1999, West Nile virus (WNv) has caused extensive bird mortality across North America, especially in American crows (Corvus brachyrhynchos) which are highly susceptible to WNv. In this study, antigenic distribution of WNv among different organs of American crows was studied, using the immunohistochemistry technique (IHC).Materials and Methods:Dead crows reported by residents were collected, transported on ice, and were necropsied for heart, lung, brain, intestine, kidney, liver, spleen, pancreas, and gonad tissues. Gross examination was performed on brain, heart, lung, liver, kidney, spleen, bursa of fabricius, gastrointestinal tract, skeletal muscle, pancreas, reproductive tract, and skin. Gross hemorrhage of brain, splenomegaly, meningoencephalitis, myocarditis, and trauma were sporadically observed in some of the infected carcasses. Formalin-fixed paraffin-embedded tissue sections were stained with IHC technique followed by counter staining with hematoxylin and eosin.Results:WNv antigen was detected in brain, spleen, heart, kidney, liver, gonads, intestine, lung, and pancreas. The spleen was found to be positive in all infected crows, followed by kidney, liver, and duodenum (95% each). Heart and pancreas were positive in 63% while brain was positive in 36.5% of the infected crows.Conclusion:More than one tissue sample is suggested to screen WNv infection using IHC technique. IHC has the advantage of correlating the visual destruction of tissue architecture with the presence of stained WNv antigen but as compared to PCR, IHC has the disadvantage of longer turnaround time, which is critical when used as a surveillance tool.

show abstract

Section: Discussioncontrasting

confidence: 57%

Antigenic distribution of west nile virus in various organs of wildly infected American crows (Corvus Brachyrhynchos)

Sandhu¹,

Sidhu

Dhillon³

2011

J Global Infect Dis

View full text Add to dashboard Cite

show abstract

“…One study reported the detection of WNV by RT-PCR in FFPE birds and horses tissues (Tewari et al, 2004). A few other studies also demonstrated the usefulness of FFPE human tissues for the detection of other RNA viruses such as Rabies virus, Sin Nombre virus, and Enterovirus by RT-PCR (Beaulieux et al, 2003;Schwarz et al, 1995;Wacharapluesadee et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Detection of West Nile virus in formalin-fixed, paraffin-embedded human tissues by RT-PCR: A useful adjunct to conventional tissue-based diagnostic methods

Bhatnagar

Guarner

Paddock

et al. 2007

Journal of Clinical Virology

View full text Add to dashboard Cite

“…Real-time PCR technology is a flexible, rapid, sensitive, specific, and quantitative method for the diagnosis of infectious disease 13 and has been used for the diagnosis of herpesviral disease. 9 Real-time PCR has been successfully used in the horse for the detection of several infectious disease agents, including: Ehrlichia spp., 18 equine infectious anemia, 5 Salmonella spp., 11 Taylorella equigenitalis, 17 West Nile virus, 25 and equine arteritis virus. 3 This article describes the development and validation of a realtime PCR diagnostic assay for detection of EHV-1 in equine nasal and blood samples and compares the performance of this assay with viral isolation.…”

Section: Introductionmentioning

confidence: 99%

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

et al. 2006

View full text Add to dashboard Cite

Equine herpesvirus-1 (EHV-1) infection is common in young horses throughout the world, resulting in respiratory disease, epidemic abortion, sporadic myelitis, or latent infections. To improve on conventional diagnostic tests for EHV-1, a real-time polymerase chain reaction (PCR) technique was developed, using primers and probes specific for the EHV-1 gB gene. Amplification efficiencies of 100% +/- 5% were obtained for DNA isolated from a plasmid, infected peripheral blood mononuclear cells (PBMCs), and nasal secretions from infected ponies. The dynamic range of the assay was 8 log10 dilutions, and the lower limit of detection was 6 DNA copies. Fifteen ponies, seronegative for EHV-1, were experimentally infected with EHV-1, and nasal samples were used to quantify shedding of virus by both virus isolation and real-time PCR analysis. Virus isolation identified nasal shedding of EHV-1 in 12/15 ponies on a total of 25 days; real-time PCR detected viral shedding in 15/15 ponies on 75 days. Viremia was quantified using PBMC DNA, subsequent to challenge infection in 3 additional ponies. Viremia was identified in 1/3 ponies on a single day by virus isolation; real-time PCR detected viremia in 3/3 ponies on 17 days. When real-time PCR was used to analyze PBMC DNA from 11 latently infected ponies (documented by nested PCR), EHV-1 was not detected. We conclude that real-time PCR is a sensitive and quantitative test for EHV-1 nasal shedding and viremia and provides a valuable tool for EHV-1 surveillance, diagnosis of clinical disease, and investigation of vaccine efficacy.

show abstract

Detection of West Nile virus using formalin fixed paraffin embedded tissues in crows and horses: quantification of viral transcripts by real-time RT-PCR

Cited by 14 publications

References 26 publications

Antigenic distribution of west nile virus in various organs of wildly infected American crows (Corvus Brachyrhynchos)

Antigenic distribution of west nile virus in various organs of wildly infected American crows (Corvus Brachyrhynchos)

Detection of West Nile virus in formalin-fixed, paraffin-embedded human tissues by RT-PCR: A useful adjunct to conventional tissue-based diagnostic methods

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

Contact Info

Product

Resources

About