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FISH Probes (fish + probe)
Selected AbstractsCorrelation between morphology and human telomerase gene amplification in bronchial brushing cells for the diagnosis of lung cancerDIAGNOSTIC CYTOPATHOLOGY, Issue 6 2010Yi-Bo Fan M.D. Abstract The aim of this study was to investigate the frequency of amplification of the human telomerase gene (TERC), as measured by fluorescence in situ hybridization (FISH), in routine liquid-based cytological preparations from bronchial brushing specimens, and to assess the associations between TERC amplification, cytological diagnosis, and cytological morphology, in order to obtain further insight into these associations. Bronchial brushings from 102 patients with lung carcinoma (52 squamous-cell carcinomas, 22 adenocarcinomas, 28 small cell lung carcinomas) and 40 patients with nonmalignant disease were used. Amplification of TERC was performed using a commercially available two-color FISH probe, and slides were prepared for the SurePath liquid-based Pap test (LPT) using the same samples. Amplification of TERC was significantly associated with histological diagnoses (P < 0.05). Patients with lung cancer, and especially those with nonsmall cell lung cancer, had significantly higher percentages of cells with amplification of TERC than did patients with nonmalignant disease (P < 0.05). Comparing the FISH and LPT results, there was no significant difference in diagnostic sensitivity between the two methods (P > 0.05). However the difference in diagnostic sensitivity of the two methods for squamous-cell carcinoma was significant (P < 0.01). FISH can be performed on bronchial brushing specimens to detect amplification of TERC. This test may be an adjunct to cytology screening, especially in squamous-cell carcinoma, and may provide an indication of the potential of individual lesions to progress. Diagn. Cytopathol. 2010. © 2009 Wiley-Liss, Inc. [source] Polar body biopsy and aneuploidy testing by simultaneous detection of six chromosomesPRENATAL DIAGNOSIS, Issue 10 2005Markus Montag Abstract Objectives To simultaneously detect six chromosomes in a single round of fluorescence in situ hybridization (FISH) during polar body diagnosis and aneuploidy testing in human in vitro fertilization (IVF) treatment. Methods A commercially available five-color FISH probe was modified by an additional chromosome probe. This kit was first tested on lymphocyte spreads and then used for polar body diagnosis (PBD) in patients with advanced maternal age and repeated implantation failure. The outcome of IVF treatment was compared with a control group. Results All six chromosomes could be simultaneously detected and easily distinguished by FISH analysis. PBD and aneuploidy testing were performed in 75 treatment cycles and compared with 126 controls. The biochemical pregnancy rate was significantly higher in the PBD group (37.1% vs 22.9%, p < 0.05) and a trend was observed for higher clinical pregnancy and implantation rates (24.22% and 14.4% vs 18.62% and 10.8%, respectively) and lower abortion rates (20% vs 31.8%) following PBD. Conclusions The simultaneous detection of six chromosomes in a single FISH round is possible and can be applied to PBD. This approach may present another step towards increasing the number of chromosomes for aneuploidy testing. Copyright © 2005 John Wiley & Sons, Ltd. [source] In situ studies of the phylogeny and physiology of filamentous bacteria with attached growthENVIRONMENTAL MICROBIOLOGY, Issue 7 2002Trine Rolighed Thomsen Summary Among the filamentous bacteria occasionally causing bulking problems in activated sludge treatment plants, three morphotypes with attached microbial growth are common, Eikelboom Type 0041, Type 1851 and Type 1701. A better knowledge of the phylogeny and physiology of these filamentous bacteria is necessary in order to develop control strategies for bulking. In this study we have used a combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR) to investigate the identity and in situ physiology of the Type 0041-morphotype and its attached bacteria in two wastewater treatment plants. Identification and enumeration of Type 0041 using group-specific 16S rRNA-targeted FISH probes revealed that approximately 15% of the filaments hybridized with a gene probe specific for the TM7 group, a recently recognized major lineage in the bacterial domain. All other filaments morphologically identified as Type 0041 only hybridized to the general bacterial EUB338-probe, indicating that they probably do not belong to commonly isolated bacterial phyla such as the Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes, for which group-specific probes were used. The phylogenetic heterogeneity of Type 0041 again highlights the inadequacy of a morphology-based classification system. Like the filaments, most of the attached microbial cells were not identified beyond their affiliation to the Bacteria using the group-specific FISH probes. However, several different bacterial phyla were represented in the identified fraction suggesting that the attached microorganisms are phylogenetically diverse. The study of the in situ physiology of Type 0041 using MAR-FISH revealed that both the filaments and the attached bacteria on Type 0041 were versatile in the use of organic substrates and electron acceptors. It was observed that all Type 0041 could consume glucose, but none of the filaments were able to consume acetate under any conditions tested, in contrast to some of the attached bacteria. No significant physiological differences were found between TM7,positive and TM7,negative Type 0041 filaments, and only minor differences were observed between the two treatment plants tested. These are the first data on the physiology of the almost entirely uncharacterized TM7 phylum and show that TM7 filamentous bacteria can uptake carbon substrates under aerobic and anaerobic conditions. [source] Oligonucleotide probes for specific detection of Giardia lamblia cysts by fluorescent in situ hybridizationJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2001M.R. Dorsch Aims:,Our study focused on the design of oligonucleotide probes and a suitable hybridization protocol that would allow rapid and specific identification of potentially viable cysts of the waterborne parasite Giardia lamblia. Methods and Results:,Comparative analysis of ribosomal RNA (rRNA) sequences of Giardia lamblia and a number of closely and more distantly related species identified six regions that appear to be specific for the G. lamblia 16S rRNA. Fluorescently labelled probes targeting these regions were produced and employed in fluorescent in situ hybridization (FISH) experiments. Two of the six probes tested successfully. Conclusions:,Our study provides the first reported probes for specific FISH detection of G. lamblia. The method depends on sufficient amounts of intact rRNA in the target organism, which is unlikely to be present in nonviable cysts that have been exposed to the environment for a prolonged period. Significance and Impact of the Study:,Currently, detection of G. lamblia cysts is largely based on immunofluorescence assays (IFA) targeting cyst wall surface antigens. These assays lack specificity and will detect species others than G. lamblia. Further, IFA will detect nonviable cysts and cyst wall fragments that do not pose a public health risk. In contrast, FISH probes allow specific detection and are likely to only detect viable, infectious cysts. [source] The yield of subtelomeric FISH analysis in the evaluation of autistic spectrum disordersAMERICAN JOURNAL OF MEDICAL GENETICS, Issue 1 2006Agatino Battaglia Abstract To assess the frequency of cryptic subtelomeric rearrangements in children and adolescents with autism spectrum disorders, blood samples were studied using a complete set of subtelomeric FISH probes in 72 children with autism spectrum disorders. All children had normal high resolution karyotype, DNA fra-X analysis, brain MRI, metabolic work-up, and physical/neirological examination. Subtelomeric analysis did not detect abnormalities in any of the subjects, suggesting the uselessness of such investigations in individuals with primary autism spectrum disorders. © 2006 Wiley-Liss, Inc. [source] |