geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Improved detection of simian immunodeficiency virus RNA by in situ hybridization in fixed tissue sections: Combined effects of temperatures for tissue fixation and probe hybridization


, : Improved detection of simian immunodeficiency virus RNA by in situ hybridization in fixed tissue sections: Combined effects of temperatures for tissue fixation and probe hybridization. Journal of Virological Methods 99(1-2): 23-32, January

In situ hybridization detection of viral RNAs in formaldehyde-fixed tissue specimens is used frequently to characterize the extent of viral replication within host tissues. The ability to determine the level of expression of viral RNAs in situ is dependent upon many factors including the extent of cross-linking during fixation, the pretreatment regimen utilized to relieve the effects of cross-linking, and the hybridization and wash protocols. In efforts to improve our ability to detect cells infected productively by simian immunodeficiency virus (SIV) in rhesus macaque tissues, the effects of unconventionally high (40degreeC) and more standard low (4degreeC) temperature fixation in 4% paraformaldehyde/phosphate buffered saline were tested empirically on in situ hybridization signals. In addition, hybridization temperatures ranging between 37 and 75degreeC were utilized to determine the optimal hybridization conditions for detection of SIV productively infected cells. Fixation conditions of 40degreeC and hybridization conditions of 50-55degreeC were identified as providing the greatest sensitivity for detecting RNA+ cells and for quantitating the signal per cell, while still allowing antigenic epitopes to be detected by immunohistochemical staining. These data indicate that the signal intensity following in situ hybridization for viral RNAs is dependent upon the combined effects of tissue fixation and in situ hybridization temperatures.

US$19.90

DOI: 10.1016/s0166-0934(01)00378-0


Other references

Konttinen, Y.T.; Tolvanen, E.; Johansson, E.; Reitamo, S.; Wegelius, O., 1982: Skin response to intradermal DNA and RNA in systemic lupus erythematosus. The local response of 18 patients with active systemic lupus erythematosus to 0.1 ml of intradermally injected 0.1% polymerized calf thymus DNA and synthetic double-stranded polyinosinic-polycytidylic acid was studied. In 14 patients positive for...

Gates, G.O., 1964: Ecology of the iguanid lizard Urosaurus graciosus in Arizona. Dissertation Abstracts, 25: 707

Ruys, J.H., 1963: Directives for vitamin D dosage.. A prophylactic dose of 600 to 800 I.U. vitamin D2 or D3 daily is recommended. When full use is made of vitaminised infant foods extra vitamin D is unnecessary.

van der Sluis, T.; Baveco, H.; Corridore, G.; Kuipers, H.; Knauer, F.; Pedroli, B.; Jochems, R.; Dirksen, J., 2003: Networks for LIFE An ecological network analysis for the brown bear (Ursus arctos) - and indicator species in Regione Abruzzo. This report gives the result of an analysis of the ecological network of Regione Abruzzo. Seven species were analysed, which can be related to 4 ecosystem types: woodland, wetland, grassland and steppe, and shrubland. The model LARCH was used to a...

Iipinge, S.N.; Pretorius, L., 2012: The delivery and quality of sexually transmitted infections treatment by private general practitioners in Windhoek Namibia. The main objective for this study was to investigate the quality of Sexually Transmitted Infections (STI) treatment and control by the private sector in Namibia. This was a cross-sectional study employing quantitative methodology using different m...

Delisle, Hélène.; Désilets, M-Claude.; Vargas, E.Ramirez.; Garrel, D., 2008: Metabolic syndrome in three ethnic groups using current definitions. According to two current definitions, the prevalence of the metabolic syndrome (MetS) among black Haitians of Montreal was <20%, 30%-36% in Algonquin Indians of Quebec, and >45% in Mexicans of Oaxaca (all aged 35-60 y). Although phenotypes w...

Poulin, B.; Wright, S.J.; Lefebvre, G.; Calderon, O., 1999: Interspecific synchrony and asynchrony in the fruiting phenologies of congeneric bird-dispersed plants in Panama. A study was conducted during 1992-94 to investigate fruiting phenologies and competition for seed dispersal in species of Miconia and Psychotria of Barro Colorado Island and Soberania National Park, central Panama. These two genera accounted for 1...

Kametani T.; Sugahara T.; Fukumoto K., 1971: Studies on total photolytic synthesis of alkaloids part 3 the products of photo pschorr reaction total synthesis of iso corydine. Tetrahedron 27(22): 5367-5374

Tsuei, D.; Fujinaga, K.; Green, M., 1972: The mechanism of viral carcinogenesis by dna mammalian viruses rna transcripts containing viral and highly reiterated cellular base sequences in adenovirus transformed cells. Proceedings of the National Academy of Sciences of the United States of America 69(2): 427-430

Green, E.L. ., 1967: Biology of the laboratory mouse Jackson Laboratory. McGraw-Hill 2nd edtn, 1966 i-xii, 1-706