Immunohistochemical test for rabies: identification of a diagnostically superior monoclonal antibody

An, Hailong; Moser, G.; Fu, ZF; Dietzschold, Bernhard; Ce, Rupprecht

doi:10.1136/vr.136.12.295

Cited by 27 publications

(14 citation statements)

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“…Este resultado difere do que já foi relatado por outros autores (Hamir & Moser 1994, Hamir et al 1995. Os anticorpos policlonais em geral apresentam alta afinidade e ampla reatividade (Ramos-Vara 2005).…”

Section: Discussionunclassified

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina

et al. 2008

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Pesq. Vet. Bras. 28(12):627-632, dezembro 2008 RESUMO.-Para a padronização da técnica de imunohistoquímica para raiva foram utilizadas cinco amostras de SNC de bovinos infectados naturalmente com o vírus da raiva usando-se um anticorpo policlonal e dois monoclonais. Para a recuperação antigênica foram avaliados os seguintes reagentes: protease XIV, proteinase K e tampão citrato pH 6,0 mantido a 100°C por 15 minutos. A detecção de antígeno rábico nas amostras foi possível com os três anticorpos utilizados. O anticorpo policlonal foi superior aos anticorpos monoclonais, demonstrando bons resultados com os três protocolos de recuperação antigênica, obtendo uma maior intensidade de marcação quando utilizado o tampão citrato e calor. A técnica de imuno-histoquímica demonstrou a presença do antígeno viral no citoplasma de neurônios na forma de agregados de grânulos ou de forma redonda ou oval, mostrando cor- One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.INDEX TERMS: Diseases of cattle, infectious diseases, diseases of the central nervous system, rabies, immunohistochemistry standardization.

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Section: Discussionunclassified

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina

et al. 2008

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“…RV minigenome (pSDI-CAT) was prepared as described previously (Conzelmann & Schnell, 1994). Monoclonal antibody to RV N (mAb 802-2) was obtained from the Centre for Disease Control and Prevention (Hamir et al, 1995) and anti-P polyclonal antibody was prepared in rabbits (Fu et al, 1994).…”

Section: Methodsmentioning

confidence: 99%

Interactions amongst rabies virus nucleoprotein, phosphoprotein and genomic RNA in virus-infected and transfected cells

Liu

Yang

et al. 2004

Journal of General Virology

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Previous in vitro studies have indicated that rabies virus (RV) phosphoprotein (P), by interacting with the nucleoprotein (N), confers the specificity of genomic RNA encapsidation by N. In this study, interactions amongst N, P and the genomic RNA in virus-infected as well as in transfected cells were studied. The results showed that when N was expressed alone, it bound non-specific RNA, particularly the N mRNA. When N and P were co-expressed, they formed N-P complexes that did not bind to non-specific RNA. When N and P were co-expressed together with (mini-)genomic RNA, N-P complexes preferentially bound the (mini-)genomic RNA. This demonstrated that RV P, by binding to N, does indeed confer specificity of genomic RNA encapsidation by N in vivo. Furthermore, the role of N phosphorylation in the N, P and RNA interactions was investigated. It was found that only N that bound to RNA was phosphorylated, while N in the N-P complex prior to RNA encapsidation was not, suggesting that RV P, by binding to nascent N, prevents the immediate phosphorylation of de novo-synthesized N. However, mutation at the phosphorylation site of N did not alter the pattern of N-P and N-RNA interactions, indicating that N phosphorylation per se does not play a direct role in N-P interaction and RNA encapsidation.

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“…Sections of FF brain were processed and examined for rabies antigen by using both anti-rabies monoclonal 6,7 and polyclonal 7,8 antibodies. Detection required a secondary reaction with either Strep-A protein 6,8 or goat antimouse antibody 7 conjugated with alkaline phosphatase.…”

Section: Subjects Materials and Methodsmentioning

confidence: 99%

“…Figure 1C shows Purkinje cells from the same case containing multiple inclusions as visualized using IHC to detect antigen. Interestingly, IHC analysis using the rabies anti-nucleocapsid MAb (802-2), which has been so effective in the detection of many strains of rabies in FF tissues, 6 did not detect this vampire bat virus. In contrast, the anti-glycoprotein MAb (2-1-7), which has detected all of the rabies strains tested thus far, 7 reacted with this vampire bat rabies antigen.…”

Section: Histopathologymentioning

confidence: 99%

Laboratory investigation of human deaths from vampire bat rabies in Peru.

Warner¹,

Zaki²,

Shieh³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. In the spring of 1996, multiple cases of an acute febrile illness resulting in several deaths in remote locations in Peru were reported to the Centers for Disease Control and Prevention (CDC). The clinical syndromes for these cases included dysphagia and encephalitis. Because bat bites were a common occurrence in the affected areas, the initial clinical diagnosis was rabies. However, rabies was discounted primarily because of reported patient recovery. Samples of brain tissue from two of the fatal cases were received at CDC for laboratory confirmation of the rabies diagnosis. An extensive array of tests on the formalin-fixed tissues confirmed the presence of both rabies viral antigen and nucleic acid. The virus was shown to be most closely related to a vampire bat rabies isolate. These results indicate the importance of maintaining rabies in the differential diagnosis of acute febrile encephalitis, particularly in areas where exposure to vampire bats may occur.

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Immunohistochemical test for rabies: identification of a diagnostically superior monoclonal antibody

Cited by 27 publications

References 0 publications

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina

Interactions amongst rabies virus nucleoprotein, phosphoprotein and genomic RNA in virus-infected and transfected cells

Laboratory investigation of human deaths from vampire bat rabies in Peru.

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