Immunogold silver staining (IGSS) was applied for the detection of porcine group A rotavirus in formalin-fixed paraffin-embedded tissue sections of small intestine. Prior to the application of IGSS, the reactivity of protein A-gold as a marker was tested with group-specific antiserum in immunogold electron microscopy. Immune aggregates were intensely and specifically labeled with the gold complex. Application of IGSS to tissue sections resulted in specific dark staining of villous enterocytes infected by group A rotavirus. This method also proved effective for the detection of rotaviral antigen in infected cultured cells. The IGSS method may be suitable for routine diagnostic detection of rotaviral infections and may have application for detection of other viral pathogens of veterinary importance.
References
1.
BendayanM: 1989, Protein A-gold and protein G-gold post-embedding immunoelectron microscopy. In: Colloidal gold: Principles, methods, and applications, ed. HayatMA, vol. 1, pp. 33–94. Academic Press, San Diego, CA.
2.
CleatorGMKlapperPEThorntonJ: 1986, Immunogold silver staining in the diagnosis of herpes encephalitis. J Neurol Neurosurg Psychiatry49:1209–1210.
3.
DanknerWMSpectorSA: 1989, Applications of immunogold-silver enhancement: Testing of monoclonal antibodies and detection of human cytomegalovirus in histologic specimens. Am J Anat185:310–313.
4.
DanknerWMSpectorSA: 1989, Detection of human cytomegalovirus by immunogold and silver enhancement. J Histotechnol12:9–13.
5.
De MeyJHackerGWDe WaeleMSpringallDR: 1986, Gold probes in light microscopy. In: Immunocytochemistry- modem methods and applications, ed. PolakJMVan NoordenS, pp. 71–88. Wright, Bristol, UK.
6.
DucatelleRCoussementWHoorensJ: 1984, Demonstration of pseudorabies viral antigen in thick and ultrathin tissue sections of young pigs using an immunogold method. Am J Vet Res45:1913–1917.
7.
FrensG: 1973, Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nat Phys Sci241:20–22.
8.
GuJDe MeyJMoeremansMPolakJM: 1981, Sequential use of the PAP and immunogold staining methods for the light microscopical double staining of tissue antigens. Regul Pept1: 365–374.
9.
HackerGW: 1989, Silver-enhanced colloidal gold for light microscopy. In: Colloidal gold: Principles, methods, and applications, ed. HayatMA, Vol. 1, pp. 297–321. Academic Press, San Diego, CA.
10.
HackerGWSpringallDRCheungA: 1986, Immunogold-silver staining and its potential use in multiple staining and histopathology. J Histochem Cytochem34:118.
11.
HackerGWSpringallDRVan NoordenS: 1985, The immunogold-silver staining method. A powerful tool in histopathology. Vichows Arch A Pathol Anat406:449–461.
12.
HainesDMChelackBJ: 1991, Technical considerations for developing enzyme immunohistochemical staining procedures on formalin-fixed paraffin-embedded tissues for diagnostic pathology. J Vet Diagn Invest3:101–112.
13.
HainesDMClarkEG: 1991, Enzyme immunohistochemical staining of formalin-fixed tissues for diagnosis in veterinary pathology. Can Vet J32:295–302.
14.
HolgateCSJacksonPCowenPNBirdCC: 1983, Immunogold-silver staining: New method of immunostaining with enhanced sensitivity. J Histochem Cytochem31:938–944.
15.
HolgateCSJacksonPLauderI: 1983, Surface membrane staining of immunoglobulins in paraffin sections of non-Hodgkin's lymphomas using immunogold-silver staining technique. J Clin Pathol36:742–746.
16.
HopleyJFADoaneFW: 1985, Development of a sensitive protein A-gold immunoelectron microscopy method for detecting viral antigens in fluid specimens. J Virol Methods12: 135–147.
17.
LindmarkRThor&-TollingKSjöquistJ: 1983, Binding of immunoglobulins to protein A and immunoglobulin levels in mammalian sera. J Immunol Methods62:1–14.
18.
MagarRRobinsonYMorinM: 1991, Identification of atypical rotaviruses in outbreaks of preweaning and postweaning diarrhea in Quebec swine herds. Can J Vet Res55:260–263.
19.
MühlpfordtH: 1982, The preparation of colloidal gold particles using tannic acid as an additional reducing agent. Experientia38:1127–1128.
20.
SpringallDRHackerGWGrimeliusLPolakJM: 1984, The potential of the immunogold-silver staining method for paraffin sections. Histochemistry81:603–608.
21.
WoodeGN: 1986, Porcine rotavirus infection. In: Diseases of swine, ed. LemanADStrawBGlockRDMengelingWLPennyRHCSchollE, 6th ed., pp. 368–382. Iowa State University Press, Ames, IA.
