Intended Use
The CEP X SpectrumOrange/Y SpectrumGreen DNA Probe Kit is
intended to detect alpha satellite sequences in the centromere region of
chromosome X and satellite III DNA at the Yq12 region of chromosome Y
in conjunction with routine diagnostic cytogenetic testing. It is indicated
for use as an adjunct to standard cytogenetic analysis for identifying
and enumerating chromosomes X and Y via FISH in interphase nuclei
and metaphase spreads obtained from bone marrow specimens in
subjects who received opposite-sex bone marrow transplantation
for chronic myelogenous leukemia (CML), acute myeloid leukemia
(AML), myeloproliferative disorder (MPD), myelodysplastic syndrome
(MDS), acute and lymphoid leukemia (ALL), or hematological disorder
not otherwise specified (HDNOS). It is not intended to be used as a
stand alone assay for test reporting; FISH results are intended to be
reported and interpreted only in conjunction with results from standard
cytogenetic analysis, performed concurrently, using the same patient
specimen. This device is not intended for use in subjects with like-sex
bone marrow transplants; with matrices other than unstimulated, cultured
bone marrow specimens; or in screening for constitutional X and Y
chromosome aneuploidies.
Limitations of the Procedure
- The CEP X/Y DNA Probe Kit has been optimized only for identifying
chromosomes in interphase nuclei or metaphase spreads from bone
marrow specimens.
- This assay identifies only the proportion of donor and recipient cells
in bone marrow specimens from recipients of opposite-sex bone
marrow transplantation. It does not distinguish between malignant
and normal cells; it is not designed to detect structural or other
chromosome abnormalities in malignant clones, which is possible
with standard cytogenetics.
- The Y chromosome is sometimes lost in bone marrow cells of
elderly males regardless of whether the specimen is from a donor,
a recipient, or collected from a patient in the post-bone marrow
transplantation period.
- It is important to have pretransplant cytogenetic results on both
donor and recipient for the following reasons: (1) There are rare
male patients who may have an unusual Y chromosome (lacking
the Yq heterochromatic region) which cannot be identified with the
CEP X/Y assay. (2) Some individuals may have target sequences
at alternate chromosome locations that hybridize to the CEP X
or Y probes. This has not been investigated for CEP X, however,
chromosome polymorphisms which hybridize with the Y probe occur
with a frequency of 1 in 2,000. Such cases may be detected
by CEP X/Y metaphase analysis and sometimes by standard
cytogenetic analysis. (3) Constitutional sex chromosome aneuploidy,
including mosaicism, present in either donor or recipient can
complicate signal enumeration and test interpretation.
- In a male donor or recipient with a 46, XY, - Y, + X karyotype, a
certain percentage of cells with XX signals will be detected by
CEP X/Y.
- If significant peripheral blood contamination is present in the bone
marrow specimen, the blood may dilute the specimen. It is important
to recognize the potential effects this dilution effect may have on the
FISH assay results; dilution of the bone marrow with blood may alter
the donor:recipient cell ratio.
- The CEP X/Y assay has been validated only for use with
unstimulated, cultured bone marrow specimens obtained from
recipients of opposite-sex BMT. It is not intended for chromosome
X and Y enumeration in other patient populations or with other test
matrices such as amniocytes, chorionic villi, fibroblasts, tumor cells,
long term cultures, among others.
- FISH assay results may not be informative if the specimen quality
and/or specimen slide preparation is inadequate.
- This device is not intended for use in subjects with like-sex bone
marrow transplants or for use in diagnostic testing or screening for
constitutional X and Y chromosome aneuploidies.
- Residual fetal cells may potentially exist in either donor or recipient
cells, however the levels at which these cells exist is likely to be
below the levels of detection of both standard cytogenetics and
FISH.
- The CEP X/Y assay has not been validated for monitoring
engraftment status.
- The clinical significance and interpretation of FISH results should
be made in conjunction with proper controls, standard cytogenetic
analysis, and within the context of the patient’s medical history and
other clinical findings.
Warnings and Precautions
- For In Vitro Diagnostic Use
- The ProbeChek Control Slides to be used with this kit are
manufactured from human cultured lymphoblast cells that have
been fixed multiple times in a solution of methanol:acetic acid (3:1).
Because it is often impossible to know which might be infectious, all
human specimens and control slides should be treated with universal
precautions. Guidelines for specimen handling are available from the
US Centers for Disease Control and Prevention.
- Hybridization conditions may be adversely affected by the use
of reagents other than those provided or recommended by
Abbott Molecular.
- Failure to follow all procedures for slide denaturation,
hybridization and signal enumeration may cause unacceptable or
erroneous results.
- Fluorophores are readily photobleached by exposure to light. To
limit this degradation, handle all solutions containing fluorophores
in reduced light. This includes all steps involved in handling the
hybridized slide. Carry out all steps which do not require light for
manipulation (incubation periods, washes, etc.) in subdued lighting to
avoid direct light projecting onto the fluorophore.
- CEP X/Y DNA Probe contains formamide, a teratogen. Avoid
contact with skin and mucous membranes. Refer to MSDS for more
information.
- The use of a calibrated thermometer is strongly recommended for
measuring temperatures of solutions, waterbaths, and incubators as
these temperatures are critical for optimum product performance.
- All hazardous materials should be disposed of according to your
institution’s guidelines for hazardous disposal.
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