Monoplex LightCycler Polymerase Chain Reaction Quantitation of the HER2 Gene for Quality Assurance of HER2/neu Testing by Immunohistochemistry and Fluorescence In Situ Hybridization

2008 ◽  
Vol 16 (6) ◽  
pp. 562-567 ◽  
Author(s):  
Lester J. Layfield ◽  
Carlynn Willmore-Payne ◽  
Gary Isom ◽  
Joseph A. Holden
Keyword(s):  
Polymerase Chain Reaction ◽  
Quality Assurance ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Chain Reaction ◽  
Her2 Gene ◽  
Polymerase Chain ◽  
Lightcycler Polymerase Chain Reaction

Combination of Cytology, Fluorescence In Situ Hybridization for Aneuploidy, and Reverse-Transcriptase Polymerase Chain Reaction for Human Mammaglobin/Mammaglobin B Expression Improves Diagnosis of Malignant Effusions

2004 ◽  
Vol 22 (3) ◽  
pp. 474-483 ◽  
Author(s):  
Michael Fiegl ◽  
Margot Haun ◽  
Anita Massoner ◽  
Jens Krugmann ◽  
Elisabeth Müller-Holzner ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Tumor Cell ◽  
Fluorescence In Situ Hybridization ◽  
Cell Detection ◽  
Malignant Cells ◽  
Chain Reaction ◽  
Tumor Cell Detection ◽  
Polymerase Chain

Purpose The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity. The aim of this study was to improve tumor cell detection in effusions by molecular approaches. Materials and Methods A total of 157 effusions from patients with tumors and 72 effusions from patients without a history or evidence of malignancy were included in this study. All effusion specimens were evaluated in parallel by cytology, fluorescence in situ hybridization (FISH) for aneuploidy, and reverse-transcriptase polymerase chain reaction (RT-PCR) for expression of human mammaglobin (hMAM) and mammaglobin B (hMAM-B). Results In effusions from patients with tumors, the sensitivities of tumor cell detection by cytology, FISH, and hMAM and hMAM-B detection were 46.2%, 53.3%, 36.4%, and 57.7%, respectively. The corresponding specificities were 94.4%, 97.0%, 87.1%, and 88.6%. Notably, a high percentage of effusions containing malignant cells were in fact transudates, indicating the necessity for molecular diagnostic work-up of transudates collected from patients with tumors. Dependent on the tumor type, the use of appropriate marker combinations improved tumor cell detection in effusions significantly. By combining all four diagnostic tests, a positive test result indicating the presence of malignancy was achieved in 81.1%, with a fairly good specificity of 70.1%. Conclusion Molecular techniques are definitely useful to detect malignancy in cytologically negative effusions. Tumor cell detection in effusions can be significantly improved by FISH and PCR techniques applying appropriate molecular markers. This finding should help to improve tumor staging, prognostic assessment, and treatment monitoring.

Orbital marginal zone lymphomas: an immunohistochemical, polymerase chain reaction, and fluorescence in situ hybridization study

2007 ◽  
Vol 38 (3) ◽  
pp. 435-442 ◽  
Author(s):  
Ginette Schiby ◽  
Sylvie Polak-Charcon ◽  
Corine Mardoukh ◽  
Kinneret Rosenblatt ◽  
Iris Goldberg ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Marginal Zone ◽  
Hybridization Study ◽  
Chain Reaction ◽  
Polymerase Chain

Isolation and Identification of Male and Female DNA on a Postcoital Condom

2000 ◽  
Vol 124 (7) ◽  
pp. 1083-1086 ◽  
Author(s):  
Stephen J. Cina ◽  
Kim A. Collins ◽  
Matthew Fitts ◽  
Mark J. Pettenati
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Sexual Intercourse ◽  
External Surface ◽  
Internal Surface ◽  
Chain Reaction ◽  
Male And Female ◽  
Polymerase Chain

Abstract Background.—Identification of male perpetrators of sexual assault may be made from cells and fluids recovered from postcoital condoms. To date, the focus has been on identifying the person who had worn the condom. Objective.—To describe a method for scientifically identifying both the male and female participants in a sex act by employing polymerase chain reaction–based technology on swabs taken from the internal and external surfaces of a condom. Fluorescence in situ hybridization may be used to screen for the presence of female cells on a condom. Methods.—Swabs were taken from the internal and external surfaces of a condom 8 hours postcoitus. DNA was isolated from each swab through standard organic extraction. Extracted DNA was amplified for 8 different genetic loci using the Promega PowerPlex kit and the sex identification amelogenin marker. Amplified samples were electrophoresed on precast sequencing gels and analyzed fluorescently using a Hitachi FMBIO 2 fluorescent scanner and software. Each DNA sample obtained from the condom was compared with male and female buccal controls. At the time of collection, air-dried slides were prepared from the swabs for subsequent multicolor fluorescence in situ hybridization using dual X- and Y-chromosome probes with 4′-6-diamidino-2-phenylindole (DAPI) counterstaining. Results.—A pure sample of female DNA was isolated from the external surface of the condom as determined by exclusive amplification of the X-chromosome–specific 212-base pair amelogenin marker. Swabs taken from the internal surface yielded DNA originating from the male participant. Identification was conclusive at 8 of 8 genetic loci. Fluorescence in situ hybridization identified pure populations of male epithelial cells from the internal surface of the condom and female cells from the external surface. Conclusions.—Cells shed from a female during sexual intercourse can be retrieved from the external surface of a condom following sexual intercourse. Fluorescence in situ hybridization can be used to screen for the presence of female cells, and positive identification of the female sexual partner can then be made using polymerase chain reaction–based methods. We suggest that swabs taken from both surfaces of a condom used during sexual assault may be used to provide information that will definitively link the victim to the suspect.

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