In situ hybridization was performed on formalin-fixed, paraffin-embedded lung sections from eight blue wildebeest (Connochaetes taurinus) calves to investigate the role of this organ in the excretion of alcelaphine herpes virus-1 (AHV-1). A digoxigenin labelled Smal fragment of AHV-1 was used as a DNA probe. Viral DNA was detected in the lungs of seven calves in which infection with AHV-1 had p...

Erythroid progenitor cells are the main target for B19 parvovirus infection. The UT7 cell line demonstrates a marked erythroid differentiation on induction by erythropoietin (EPO) (UT7/EPO cells) and therefore appears to be a potential target for B19 parvovirus. We aimed to evaluate the presence and localization of B19 nucleic acids in UT7/EPO cells by in situ hybridization. Three digoxigenin-l...

The present study describes a colony hybridization setup for identification of enterotoxigenic Escherichia coli obviating the need for advanced equipment and radioactive isotopes. With a modest laboratory arrangement, polynucleotide gene probes were produced in large quantities. The probes were labeled with digoxigenin and, after hybridization, detected with an antidigoxigenin alkaline phosphat...

Using cultured mouse fibroblast L929 cells, this study demonstrated the hemolytic and cytotoxic activities and induction of apoptosis in cells infected with Orientia tsutsugamushi. Low levels of hemolytic activity were detected using heavily infected cells. No hemolysin or cytotoxin were detected in the infected culture fluid regardless of the pathogenicity of the O. tsutsugamushi strains in mi...

A PCR assay for the diagnosis of infection produced by Candida sp. was developed. The primers, designated 520 and 522, were selected from highly-conserved areas of the small subunit (ssu) 18S rRNA gene of Candida spp. To check the value of the results a Candida albicans oligonucleotide probe, digoxigenin-labeled, and a general Candida probe were used in hybridization experiments with the amplif...

This paper describes a method for non-radioactive in situ hybridization providing subcellular localization of mRNA in 3 microns cryosections. We used two alternative colorimetric reactions to detect digoxigenin-labeled cRNA probes: alkaline phosphatase and HRP (horseradish peroxidase). With some probes the signal with the alkaline phosphatase reaction was intense, and diffusion of the reaction ...

We describe a method for the in situ detection of specific splicing variants. The method is based on the use of antisense oligonucleotides designed to span splice junctions labelled with digoxigenin by terminal transferase tailing. We find that the spatial patterns of Ubx splicing variants Ia and IIa are similar in early embryos, but differ in late embryos. Variant IVa is only detected in the C...

Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as approximately 14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5'-digoxigenin-endlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)...

=Abstract=ln situ hybridization using digoxigenin labeled HPV(human papillornaviruses) probes was tried in order to detect the presence of HPV DNA in 27 cases of single and multiple laryngeal papilloma. There were HPV6/11 DNA signals in 17 out of 22 cases of multiple laryngeal papilloma and in all five cases of single laryngeal papilloma. The specific signals were seen focally in the nuclei of ...

We have explored a simplified system without purification for systematic evolution of ligands by exponential enrichment (SELEX) against Vibrio alginolyticus to select single-stranded DNA ligands (aptamers) from a random 82-nt library. The DNA content was quantified by agarose gel electrophoresis and follow-up image analyses with the BandScan 5.0 software. Anti-digoxigenin/HRP system was used to...