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Resources for the Laboratory Professional
Sample Images of FISH
Results may vary.
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Normal chromosome number for 18, X and Y A cell containing one green X signal and one orange Y signal, indicative of the normal XY and two aqua signals, indicative of the normal two copies of chromosome 18. |
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Trisomy 18 and normal chromosome number for X and Y A cell containing one green X signal and one orange Y signal indicative of the normal XY and three aqua signals, indicative of the trisomy 18. |
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Normal chromosome number for 13 and 21 A cell containing two orange 21 probe signals indicative of the normal chromosome 21 number and two green signals indicating the normal chromosome 13 number. |
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Trisomy 21 and normal chromosome number for 13 A cell containing two green 13 probe signals indicative of the normal chromosome 13 number and three orange chromosome 21 probe signals indicative of trisomy 21. |
Laboratory Procedure for Uncultured Amniotic Fluid (3-stage Protocol)
For more information, please consult the AneuVysion product insert.
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1. Prepare Slides Treat a 2 to 5 mL specimen of amniotic fluid with hypotonic solution to expand the cells. Add fixative. Place the cells on a glass slide creating 2 hybridization areas. |
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2. Bind Probe To Target Place the slide in a heated formamide solution to denature the cellular DNA. Apply DNA probes (the LSI 13 and 21 probe and the combinded CEP 18, X and Y probes) to their target areas on the slide. |
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Allow probes to hybridize to the complementary DNA for 6-24 hours. Wash slide to remove unbound probe. |
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3. Analyze Apply counterstain, view and enumerate probe signals using a fluorescence microscope. |
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A. CEP 18,X,Y: One green signal (CEP X) one orange signal (CEP Y) and three aqua signals (CEP 18) indicate one copy of the X and Y chromosome and three copies of the 18 chromosome (XY,+18).
B. LSI 13,21: Two green signals (LSI 13) and two orange signals (LSI 21) indicate two copies of chromosome 13 and two copies of chromosome 21. |
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Kit Components
Our easy-to-use kits are an easy and economical way to reproducibly perform standardized assays. Each kit contains the essential components needed to perform standardized assays (22 x 22 mm area). Additional reagent purchases are minimal.
Fluorescence Microscope Filter Sets - See table for technical information on recommended filter sets. Contact Vysis' Technical Service Department for additional information on filter set requirements.
Control Slides
ProbeChek Control Slides
Control slides are required to monitor the assay performance and assess the accuracy of the signal enumeration.
Prenatal Positve: 05J36-005
Prenatal Negative: 05J39-005
Reagents
20X SSC
For preparation of denaturation and wash solutions.
NP-40
For preparation of wash solutions.
DAPI II
For counterstaining of cells
Recommended Optical Sets
Technical information on recommended filter sets.
(Viewed with fluorescence microscope and 100-watt lamp source.)
Although the SpectrumGreen LSI 13 and SpectrumOrange LSI 21 are visible using the triple bandpass filter set, the single bandpass filter sets may provide a better view in some specimens.
References
| Study | Summary |
Download
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Rapid prenatal diagnosis of trisomy 21 in 5,049 consecutive uncultured amniotic fluid samples by fluorescence in situ hybridization (FISH) |
This was a retrospective study on the results of interphase fluorescence in situ hybridization (FISH), performed routinely for chromosome 21 and on ultrasonographic indications for chromosomes 13, 18, X and Y in a series of 5,049 amniotic fluid samples. |
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| Prenatal diagnosis using interphase fluorescence in situ hybridization (FISH): 2-year multi-center retrospective study and review of the literature | Data is presented from the clinical trial that led to regulatory clearance for AneuVysion (1379 pregnancies) and from retrospective case review on 5197 new pregnancies. | Full PDF |
| Rapid Prenatal Diagnosis of Aneuploidies in Uncultured Amniocytes by Fluroescence in situ Hybridization | A new method in prenatal diagnostics allows the identification of certain numeric chromosomal aneuploidies in amniotic cells within 24 hours in contrast to conventional methods which take 1-3 weeks. | Full PDF |
| Rapid Prenatal Diagnostics in the Interphase Nucleus : Procedure and Cut-off Rates | Results of a rapid prenatal aneuploidy screening in uncultured amniocytes using FISH are available within 24 hr. Here we present our cut-off rates for and the way in which we handle the AneuVysion kit (ABBOTT/Vysis), based on the experience of 1200 studied cases. In summary, the method is reliable, and the risk for misdiagnosis is low (_0.4%). | Full PDF |
| Automated Microscopy of Amniotic Fluid Cells: Detection of FISH Signals Using the FastFISH® Imaging System | FISH (fluorescence in situ hybridization) analysis is a valuable adjunct to cytogenetics that provides a rapid screen for common abnormalities. A fully automated system for FISH analysis could improve laboratory efficiency and potentially reduce errors and costs. | Free PDF unavailable. Available for purchase. |
Ordering AneuVysion
Sources
1.Klinger K, Landes G, Shook D et al. Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization (FISH). Am J Hum Genet 1992; 51: 55-65.
2.Evans MI, Ebrahim SA, Berry SM et al. Fluorescent in situ hybridization utilization for high-risk prenatal diagnosis: a trade-off among speed, expense, and inherent limitations of chromosome-specific probes. Am J Obstet Gynecol 1991; 1055-1057.
3.Lewin P, Kleinfigner P, Bazin A et al. Defining the efficiency of fluorescence in situ hybridization on uncultured amniocytes on a retrospective cohort of 27,407 prenatal diagnoses. Prenat Diagn 2000; 20: 1-6.
4.American College of Medical Genetics (ACMG). Technical and clinical assessment of fluorescence in situ hybridization: an ACMG/ASHG position statement. I. Technical considerations. Genetics in Medicine 2000; 26:356-61.
5.Tepperberg J, Pettenati MJ, Rao PN et al. Prenatal diagnosis using interphase fluorescence in situ hybridization (FISH), 2-year multi-center retrospective study and review of the literature. Prenat Diag 2001; 21: 293-301.
