| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
From the Division of Pathology,
*
City of Hope National Medical Center, Duarte, California; the Department of Pathology,
Oregon Health Sciences University, Portland, Oregon; the Department of Pathology and Laboratory Medicine,
University of Pennsylvania, Philadelphia, Pennsylvania; and the Department of Cellular Pathology,
Armed Forces Institute of Pathology, Washington, DC
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
chain gene rearrangements. Of 14 laboratories
performing TCRß Southern blot analysis, there was
complete agreement in 10 of 14 cases, with some false negative
results obtained in 4 cases. No false positive results were obtained by
Southern blot analysis. TCRß PCR analysis was only
performed by two laboratories, and only 47.1% of positive
samples were detected. Twenty-one laboratory results were obtained for
TCR PCR. This method showed an overall detection rate of
77.9% for T cell gene rearrangements with a 4.1% false positive
rate, as compared to both TCR Southern blot
analysis results and immunophenotyping. The detection rate for
TCR PCR, however, significantly differed
when extracted DNA samples from frozen tissue were compared to
paraffin-embedded tissue (85.4% versus 65.9%;
P = 0.0005). Significant differences in true
positive results were obtained when laboratories using primers directed
against multiple TCR variable regions (V1–8 plus one to
three other primer sets) were compared to laboratories that used only a
single set of TCR primers directed against the V1–8
(P < 0.0001). Other technical factors
significantly affecting results were also identified. These findings
provide useful data on the current state of diagnostic TCR
testing, highlight the risk of false negative results for TCR
testing directed against only portions of the TCR
gene, and identify limitations of testing of paraffin-embedded
tissues in some laboratories. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The four T cell receptor genes undergo variable-diversity-joining (VDJ)
region or variable-joining (VJ) region rearrangements as part of normal
T cell development, similar to the immunoglobulin heavy chain and light
chain gene rearrangements of B cell development.3, 4
The T
cell receptor 
locus (TCR) at chromosome region 14q11
is the first to rearrange, followed by TCR at 7q15,
TCRß at 7q34, and, finally, TCR
at 14q11.
Approximately 95% of circulating T cells undergo all four
rearrangements, but a small percentage of cells (/ T cells) only
undergo rearrangement of the first two genes. In the past, the most
commonly used methods of detecting T cell receptor gene rearrangements
used Southern blot analysis directed against the constant or joining
regions of the TCRß gene.5
Southern blot
methods, however, are time-consuming, labor intensive, require a
relatively large amount of fresh or frozen tissue, and require at least
5 to 10% clonal cells in the sample for detection. More recently,
polymerase chain reaction (PCR)-based methods of detection of gene
rearrangements have been used.2
These methods target
rearrangements of either the TCRß gene or the TCR gene;
but because the TCR variable region is less complex than
the TCRß variable region, many laboratories prefer
TCR testing. These methods are more rapid, require
smaller amounts of tissue, may be used in paraffin-embedded tissues and
can detect a smaller percentage of clonal cells than Southern blot
analysis.
To evaluate the utility and methodology used in laboratories performing T cell receptor clonality studies, the authors circulated a total of 29 samples of paraffin-embedded tissue and DNA extracted from frozen tissue from B and T cell lymphomas or leukemias to 21 diagnostic laboratories for PCR testing. Thirteen laboratories also performed Southern blot analysis on 14 of the samples, resulting in a total of 24 participating laboratories. The results of this sample exchange provide more information about the different testing methodologies used in diagnostic laboratories and their use in fresh/frozen and paraffin-embedded samples.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Frozen cells and available corresponding paraffin-embedded tissue of archived lymphoma and leukemia samples originally diagnosed at the Oregon Health Sciences University Department of Pathology were retrieved for use in this study. Lineage was assigned based on prior immunophenotyping studies in conjunction with morphological evaluation. Cases were classified according to the Revised European-American Classification of lymphoid neoplasms and the proposed World Health Organization Classification.1, 6 Extracted DNA from 16 lymphoma/leukemia samples (9 B-lineage: 7 follicular lymphoma, 2 chronic lymphocytic leukemia/small lymphocytic lymphoma; 7 T-lineage: 4 peripheral T cell lymphoma, 1 mycosis fungoides/Sézary syndrome, 2 lymphoblastic lymphoma) was aliquoted and distributed to each participating laboratory. For DNA extraction, frozen cells were thawed, pelleted, and resuspended in an equal volume of lysis buffer, consisting of 50M Tris-HCL, pH 8.0, 100 mmol/L EDTA, supplemented with 0.9 mg/ml proteinase K and 0.5% SDS. The lysis mix was incubated for 16 hours at 55°C and extracted twice with an equal volume of phenol and once with chloroform/isoamylalcohol at a 24:1 ratio. DNA was precipitated with 0.1 volume of 7.5 mmol/L NH4OAc and 2.5 volumes of 100% ethanol, washed in 70% ethanol, and air-dried. The pellet was resuspended in TE (10 mmol/L Tris-HCl, pH 8.0, 1 mmol/L EDTA) and incubated for 1 hour at 55°C. If DNA was not completely dissolved, the TE incubation process was repeated as necessary. DNA concentration was determined and dilutions were made with TE to a final concentration of 0.1 µg/µl. 500 µl (50 µg DNA), or 100 µl (10 µg DNA) were aliquoted for distribution to the participating laboratories.
Samples from 13 formalin-fixed paraffin-embedded lymphoma/leukemia specimens were also distributed. These included 11 paraffin samples that corresponded to frozen cell specimens (5 B-lineage: 4 follicular lymphoma and 1 chronic lymphocytic leukemia/small lymphocytic lymphoma; 6 T-lineage: 4 peripheral T cell lymphoma, 1 lymphoblastic lymphoma and 1 mycosis fungoides, Sézary syndrome), and two additional paraffin-embedded specimens (1 B-lineage: mantle cell lymphoma; 1 T-lineage: peripheral T cell lymphoma). For paraffin-embedded tissues, four or five 10-µm sections of each block were distributed in tubes to each participating laboratory and DNA extraction was performed in the individual laboratories. Laboratories were not aware that that some frozen and paraffin samples were from the same tumor.
Results were compared between laboratories with the exception of
Southern blot analysis for rearrangements of the T cell receptor
chain gene (TCR), which was performed in one laboratory
for comparison to the polymerase chain reaction testing for
rearrangements of this locus. For TCR Southern blot, 16
µg of extracted DNA was digested with HindIII and
EcoRI for 3 hours. The digested DNA was loaded on a 0.7%
agarose gel and run at 34 V for 16 hours at room temperature. After
electrophoresis, the DNA was transferred overnight with 0.4 N NaOH onto
a nylon membrane. The blots were hybridized with H60 probe (ATCC 59585,
Rockville, MD).7
H60 probe was labeled with P-32
nucleotides by use of a random primer labeling kit (Amersham Pharmacia,
Arlington Heights, IL). The blots were washed with 0.1X SSC and 0.1%
SDS, and exposed to X-ray films for 5 days. The presence of additional
bands or loss of bands when compared to a germline control was
considered evidence of a TCR gene rearrangement.
The submitting laboratory, using their own established guidelines, interpreted the test results. Specific guidelines for interpretation were not proposed by the exchange.
Samples were accompanied by a technical questionnaire related to the individual test performed and laboratories were requested to return the completed questionnaire, a summary of results and copies of diagnostic radiographs and/or gels used for interpreting results.
2 analysis of probability was performed using
GB-STAT version 7.0 (Dynamic Microsystems, Inc., Silver Spring, MD).
Probability (P) values of 0.05 or less were
considered statistically significant.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
PCR by two different
methods and was counted twice to give a total of 25 laboratory results.
Thirteen laboratories performed Southern blot analysis for TCR gene
rearrangements on some or all of the extracted DNA specimens, two
performed TCRß PCR, and 21 performed TCR
PCR.
Technical Questionnaire
The technical questionnaire for TCR Southern blot analysis
indicated that laboratories routinely used two to five different
restriction enzymes with the majority of laboratories(9 of 13) using
BamHI, EcoRI, and HindIII. Of the 11
laboratories responding, three required only one rearranged band for a
positive result and the remaining eight required two rearranged bands
before interpreting a case as rearranged. Seven laboratories used
isotopic probes labeled in their own laboratory and seven used
non-isotopic methods. Six laboratories used
JßI-II probes only, four used CTß probes only
and three used both.
The technical questionnaire on TCR PCR indicated that
paraffin section extraction methods varied from organic in 7 (33%),
inorganic in 7 (33%), and crude lysate in 5 (24%) laboratories. No
information was received from two (10%) of the participating
laboratories. Twelve centers (57%) used a "hot start" PCR approach
and 8 (38%) used standard PCR. One center gave no information
regarding the PCR approach. All 20 responding laboratories used a
non-nested PCR method, but the TCR variable regions
covered by the primers varied. Nine laboratories (43%) used a
combination of primers directed against all 11 V regions (V 1–8,
9, 10, 11) as well as a multiplex of J primers (Group
1);8, 9, 10, 11
4 centers (19%) used a J multiplex with
primers directed against V 1–8, 10 and 11 (Group
2);12, 13, 14, 15
1 laboratory (5%) used primers directed against
V 1–8, and 9 with a multiplex of J primers (Group
3);8
and, 5 (25%) used a single set of primers directed
against V 1–8 and J (Group 4).16, 17, 18, 19
The V
regions covered by the PCR primers were not available for 2
laboratories. The J primers used were variable within the groups. In
Group 1, two laboratories used a multiplex of J1, JP, JP1 and JP2; 1
laboratory each used the following combinations: J2, JP, JP1 and JP2;
J1 and J2; J2 and JP2; J1/2 and JP; J1/2, JP, JP1 and JP2; and J1/2 and
JP1. Two laboratories in Group 2 used multiplex of J2 and JP2, while
the other 2 laboratories in this group used the combinations of J2, JP
and JP2, and J2, J1/2 and JP2. The single laboratory in Group 3
reported use of a multiplex of J2, JP, and JP1. Three of the
laboratories in Group 4 reported use of a single set of J2 primers, one
used a single set of J1/2 primers and specifics of the single primer
set used were not provided by one laboratory.
PCR products were analyzed on polyacrylamide gel in 14 laboratories (66%), by capillary electrophoresis in 3 (14%) and by agarose, MetaPhor (Cambrex, East Rutherford, NJ), and denaturing gradient gel electrophoresis (DGGE) in 1 laboratory each. The method of analysis was not reported for one laboratory. Denaturing conditions (heteroduplex) were used in 6 of the 20 laboratories with data. Ethidium bromide staining was used by 12 of 19 reporting labs, by silver staining in 2 labs, by SYBR green staining in 2 laboratories, and by Gelstar, incorporated radionucleotide and chemiluminescence in 1 laboratory each.
Types of controls included patient samples (9 laboratories; 43%) and
cell lines (10 laboratories; 47%). Types of controls were not reported
for 2 laboratories (10%). Fifteen laboratories gave information on the
type of sensitivity controls that they used, with diluted cells used by
3 and diluted DNA by 12. The range of sensitivities predicted by the
laboratories was 0.001 to 10% for frozen samples and 0.01 to 10% for
paraffin samples. Laboratories predicted that their TCR
methodology could detect 75 to 95% of clonal T cell rearrangements
with a mean of 86%.
TCR Southern Blot
Southern blot analysis showed rearrangement of the
TCR gene in all cases of T cell lymphoma and none of the
B cell neoplasms.
Exchange Results
The Southern blot results are summarized in Table 1
. Although all laboratories that performed Southern blot analysis for
TCR gene rearrangements did not test every specimen, presumably due to
a lack of sufficient DNA for all tests covered by the sample exchange,
there was concordance among all testing laboratories on 10 of 14
samples. The complete concordance was on four B cell neoplasms (all
TCRß germline) and six T cell neoplasms (all
TCRß rearranged). Differences in results in four cases
included one follicular lymphoma (eight laboratories TCRß rearranged
and two germline), one mycosis fungoides/Sézary syndrome (11
laboratories TCRß rearranged, one germline), and two
peripheral T cell lymphomas (one case with 4 laboratories
TCRß rearranged and three germline; one case with 12
laboratories TCRß rearranged and one germline). For the 5
laboratories with a discordant result, no trend related to probe type
was identified when compared to the overall results. Two used CTß
probes and three used JßI-II. Two of the
laboratories with discordant results used isotope-labeled probes and 3
used non-isotopic probes. No discordant results were reported from the
3 laboratories that used both CTß and JßI-II
probes, although 1 of those laboratories only tested 3 of the 14
available samples. Based on the consensus of the participating
laboratories of no TCRß rearrangements detected 6 of 7 of
the B cell neoplasms, and apparent dual T and B cell rearrangements in
one B cell lymphoma, the results of the survey were interpreted as
representing no false positive results for TCRß Southern
blot analysis.
|
2summarizes the results from these laboratories, compared to the
consensus Southern blot results for TCRß (including one
TCRß-positive B cell neoplasm). Only 47.1% of expected
TCRß rearrangements were detected by this method with a
15% false positive rate.
Polymerase chain reaction testing for TCR was performed
in 21 laboratories, and the results are summarized in Tables 2
and 3
. Overall, these tests detected 77.9% of expected T cell gene
rearrangements. There was a significant decrease in the ability to
detect the gene rearrangement in paraffin-embedded tissue, ranging from
85.4% of extracted DNA samples and 65.9% of paraffin samples
(P = 0.0005). A 4.1% false positive rate was
detected, overall, and no difference was seen between paraffin and
extracted DNA samples. Almost 22% of samples, however, were not
tested, presumably due to the lack of amplification of an internal
control gene. A lack of testing was significantly higher for the
paraffin embedded samples (7.1% of extracted DNA samples
versus 39.6% for paraffin samples; P <
0.0001).
|
|
results were further analyzed by paraffin
extraction methods, PCR method (hot start versus standard),
DNA analysis method (capillary electrophoresis versus
polyacrylamide gel and denaturing versus non-denaturing gel
analysis) and TCR variable and joining regions covered by
the PCR primers. These results are summarized in Tables 4
5
6
7
8
. The use of inorganic extraction methods of paraffin tissue resulted
in a slight, but significant (P = 0.0059)
increase in false positive results, when compared to organic and crude
lysate methods (Table 4)
. Although a significant difference in results
was not observed when hot start PCR was compared to standard PCR, more
cases could not be analyzed due to lack of amplifiable DNA when
standard PCR methods were used (P = 0.0053)
(Table 5)
. When capillary electrophoresis was compared to polyacrylamide gel
analysis of DNA products, a slight increase in paraffin false positive
results was observed with capillary electrophoresis (2/12 or 16.7%
versus 1/53 or 1.9%; P = 0.0276) (Table 6)
. However, significantly fewer cases could be analyzed using
polyacrylamide gels (20.2% polyacrylamide gel paraffin cases not
tested versus 10.3% for capillary electrophoresis; P =
0.0316). Although there was a slight increase in extracted frozen
samples not tested when non-denaturing conditions were compared to
denaturing gels (P = 0.0257), no other
significant differences between these methods were identified (Table 7)
.
|
|
|
|
|
PCR
(Table 8)
, failed to identify a significant difference in false positive results
with any primer group. However, primers strategies devised to amplify
V 1–8, and 9 (with or without V 10 or 11) with a multiplex of
J primers (Groups 1–3), had significantly higher true positive rate
for extracted frozen cell DNA (P = 0.0082),
paraffin samples (P = 0.0027) and all samples
(P < 0.0001) when compared to Group 4, which
used a single a single set of primers to detect V 1–8 and J. |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Few conclusions can be drawn from the results of the TCRß polymerase chain reaction sample exchange due to the small number of laboratories performing this test. The results suggest, however, that the methods used by participants in this exchange for detecting TCRß by PCR fail to detect a significant number of cases, with just over half of the T cell neoplasms detected in this study.
More information can be drawn from the TCR PCR sample
exchange. Testing for rearrangement of the TCR gene by
PCR analysis has been increasingly popular in diagnostic laboratories.
This gene is rearranged earlier in T cell development than the
TCR and ß genes, and therefore TCR gene
rearrangements are detectable in virtually all T cell
neoplasms.4, 10, 11
In addition, the number of variable and
joining regions of TCR is less than for
TCRß, with only four variable region families made up of a
total of 11 variable loci and 5 joining region loci. PCR testing
directed at the four variable region families with a multiplex of
joining region primers can theoretically detect all TCR
rearrangements.2
The results of the exchange highlight the
extreme variability in the methods used for this test. This test is
performed well in many laboratories, but several laboratories had
suboptimal results when testing paraffin embedded tissues. All
laboratories estimated in the technical survey that they could detect
at least 75% of T cell neoplasms, with many reporting a 95% expected
detection. Despite this, the overall detection rate in paraffin tissues
was 65.9% in this study. This suggests that a significant number of
laboratories have not actually compared their results for paraffin and
fresh/frozen samples and may not realize the potential risk of false
negative results on paraffin tissues. Difficulty in extracting
sufficient DNA for analysis from the paraffin tissue is the most likely
cause for this drop in detection. This difficulty is also suggested by
the significantly higher number of paraffin samples that were not
tested in the exchange, presumably due the inability to extract
sufficient DNA for analysis and the resulting failure to amplify an
internal control gene. An increase in the inability to test samples was
also related to use of standard PCR (in comparison to hot start PCR),
use of non-denaturing gel conditions, and use of polyacrylamide gels
(compared to capillary electrophoresis). These results suggest that
improved extraction methods, hot start PCR methods and more sensitive
detection methods may reduce the number of false negative results for
this test.
False positive results for TCR PCR were not common in the
sample exchange, occurring at a rate of 3.6% for extracted frozen
tissue DNA, 5.4% for paraffin samples and 4.1% overall. One parameter
identified to be related to an increase in false positives was use of
inorganic extraction methods on paraffin embedded tissues. Although
these methods yielded a slight increase in true positive results
(76.5%) when compared to organic (61.8%) and crude lysate (42.3%)
methods, they also resulted in a slight, but statistically significant
increase in false positive results. The use of capillary
electrophoresis was also associated with an increase in false positive
results when compared to the use of polyacrylamide gel electrophoresis.
Therefore, although the use of this newer methodology appears to
increase the detection of rearrangements, it may also introduce an
increase risk for false positive results. The increase in false
positive results with capillary electrophoresis could be related to
over-interpretation of results with this very sensitive methodology. As
more laboratories begin using this method of detecting
TCR rearrangements, the potential for false positive
results should be recognized and stringent interpretative criteria
should be developed to reduce this possibility.
An increase in false positive TCR PCR has been previously
reported with the use of primers directed against V9. Some
circulating / T cells reportedly undergo normal polyclonal gene
rearrangements with little junctional diversity. These so-called
canonical TCR rearrangements do not demonstrate nucleotide additions
and therefore may produce polyclonal rearrangements that are very
similar or identical in size. This phenomenon is reported to occur more
frequently with rearrangements involving V9 and
JP.21
No such increase in false positive results was
identified in this study in Groups 1–3, which included primers
directed against the V9 region and all of the J regions of the
TCR gene.
The primer strategy used to detect TCR rearrangements by
PCR analysis was a highly important factor in the ability of
laboratories to detect rearrangements on a consistent basis.
Laboratories that used a single set of primers directed against
consensus regions of J and V1–8 had the lowest rate of
detection. The addition of more primers sets, including primers to
cover all J regions and some or all of the other V regions
(V9, 10, 11), significantly increased the ability to detect
TCR gene rearrangements without increasing the rate of
false positive results. The cases tested in this exchange were randomly
chosen without consideration of V region rearrangements.
Recently, a multicenter trial of German, Austrian, and Swiss
investigators reported their results for TCR PCR
testing.22
In that study, 26 laboratories studied six
paraffin samples, including four T cell neoplasms and two reactive
hyperplasias. Overall, an average of 90% of TCR results
was considered "correct" in that study, with correct results
ranging from 69% to 100%. Further details of the results were limited
in that report. Although the German-Austrian-Swiss study results
suggest a higher detection rate than the results of the current study,
the small number samples and the lack of detailed results for the
former study limits the comparison of the two multicenter studies.
Recent attempts to standardize PCR testing have also been published.
European laboratories have developed standardized primers and protocols
for the detection of minimal residual disease in acute lymphoblastic
leukemia, which include testing for TCR.23
The reproducibility of this approach, however, has not yet been
reported.
In summary, this sample exchange for T cell receptor testing provides a
large amount of data, but several key points can be made: 1) Southern
blot analysis for TCRß chain gene rearrangements remains a
reliable and reproducible test with minimal false positive results. 2)
TCR PCR testing is also a reliable and reproducible test
that is performed well in many laboratories, especially on fresh or
frozen tissues. 3) Paraffin sample TCR PCR testing is
less reliable than fresh or frozen tissue testing in most laboratories,
and the risk for false negative results for paraffin tissues should be
recognized. 4) A high number of samples, particularly paraffin tissues,
could not be reliably tested due to a lack of amplifiable DNA. This
appears to be related to a variety of technical factors. 5) Single
primer set strategies that are only directed against a consensus of the
V1–8 and J regions of TCR are suboptimal for the
detection of many clonal T cell neoplasm samples.
Based on the findings of this exchange, three recommendations can be
made. First, the authors continue to support previously published
criteria for the interpretation of Southern blot studies that require
the identification of two rearrangements (either two rearrangements in
one digest or one rearrangement in two different
digests).24
Second, PCR testing for TCR gene
rearrangements should employ primers directed against all areas of the
TCR V and J regions to minimize false negative results.
Finally, laboratories that wish to perform testing in paraffin-embedded
tissues should separately verify their methodology on that tissue type.
|
Acknowledgments |
|---|
Southern blot testing.
Participating laboratories were: Armed Forces Institute of Pathology,
Washington, D.C.; Barnes-Jewish Hospital, Washington University, St.
Louis, MO; City of Hope National Medical Center, Duarte, CA; DAKO
Corporation, Carpinteria, CA; Evanston Hospital, Northwestern
University, Evanston, IL; Georgetown University Medical Center,
Washington, D.C.; Loyola University Medical Center, Maywood, IL;
Marshfield Laboratories, Marshfield, WI; Mayo Clinic, Rochester, MN;
Medical University of South Carolina, Charleston, SC; Oregon Health
Sciences University, Portland, OR; Roswell Park Cancer Institute,
Buffalo, NY; Rush Medical Center, Chicago, IL; St. Barnabas Medical
Center, Livingston, NJ; SUNY Hospital of Stony Brook, Stony Brook, NY;
SUNY University Hospital of Syracuse, Syracuse, NY; University of
Florida, Gainesville, FL; University of Missouri, Columbia, MO;
University of Pennsylvania, Philadelphia, PA; University of Texas
Health Science Center, San Antonio, TX; University of Texas Medical
Branch, Galveston, TX; University of Texas Southwestern, Dallas, TX;
William Beaumont Hospital, Royal Oak, MI; and University of New Mexico
Health Sciences Center, Albuquerque, NM. |
Footnotes |
|---|
Results were preliminarily presented at the AMP annual meeting in 1999.
Accepted for publication August 31, 2001.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-gene by a site-specific recombination. Nature 1988, 335:170-174[Medline]
/ lineage relationship within a consecutive series of human precursor T-cell neoplasms. Blood 1989, 74:2508-2518 gene rearrangement in the diagnosis of cutaneous lymphocytic infiltrates. Diagn Mol Pathol 1994, 3:275-282[Medline]
gene rearrangements by denaturing gradient gel electrophoresis of GC-clamped polymerase chain reaction products: correlation with tumor-specific sequences. Am J Pathol 1995, 146:46-55[Abstract]
locus in peripheral T-cell lymphomas: analysis by polymerase chain reaction and denaturing gradient gel electrophoresis. J Pathol 1996, 178:303-310[Medline]
chain gene. Diagn Mol Pathol 1992, 1:173-179[Medline]
This article has been cited by other articles:
 |
|
 |
|
  K. P. Patel, Q. Pan, Y. Wang, R. W. Maitta, J. Du, X. Xue, J. Lin, and H. Ratech Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-{gamma} Gene Rearrangements J. Mol. Diagn., March 1, 2010; 12(2): 226 - 237. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. L. Salisbury, S. D. Budnick, and S. Li T-Cell Receptor Gene Rearrangement and CD30 Immunoreactivity in Traumatic Ulcerative Granuloma With Stromal Eosinophilia of the Oral Cavity Am J Clin Pathol, November 1, 2009; 132(5): 722 - 727. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. T. Tan, K. Seo, R. A. Warnke, and D. A. Arber The Frequency of Immunoglobulin Heavy Chain Gene and T-Cell Receptor {gamma}-Chain Gene Rearrangements and Epstein-Barr Virus in ALK+ and ALK- Anaplastic Large Cell Lymphoma and Other Peripheral T-Cell Lymphomas J. Mol. Diagn., November 1, 2008; 10(6): 502 - 512. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Bagg Malleable Immunoglobulin Genes and Hematopathology - The Good, the Bad, and the Ugly: A Paper from the 2007 William Beaumont Hospital Symposium on Molecular Pathology J. Mol. Diagn., September 1, 2008; 10(5): 396 - 410. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. T. Tan, R. A. Warnke, and D. A. Arber The Frequency of B- and T-Cell Gene Rearrangements and Epstein-Barr Virus in T-Cell Lymphomas: A Comparison Between Angioimmunoblastic T-Cell Lymphoma and Peripheral T-Cell Lymphoma, Unspecified With and Without Associated B-Cell Proliferations J. Mol. Diagn., September 1, 2006; 8(4): 466 - 475. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Sandberg, E. J. van Gastel-Mol, B. Verhaaf, K. H. Lam, J. J.M. van Dongen, and A. W. Langerak BIOMED-2 Multiplex Immunoglobulin/T-Cell Receptor Polymerase Chain Reaction Protocols Can Reliably Replace Southern Blot Analysis in Routine Clonality Diagnostics J. Mol. Diagn., October 1, 2005; 7(4): 495 - 503. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. C. Lawnicki, R. J. Rubocki, W. C. Chan, D. M. Lytle, and T. C. Greiner The Distribution of Gene Segments in T-Cell Receptor {gamma} Gene Rearrangements Demonstrates the Need for Multiple Primer Sets J. Mol. Diagn., May 1, 2003; 5(2): 82 - 87. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. C. Greiner and R. J. Rubocki Effectiveness of Capillary Electrophoresis Using Fluorescent-Labeled Primers in Detecting T-Cell Receptor {gamma} Gene Rearrangements J. Mol. Diagn., August 1, 2002; 4(3): 137 - 143. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Bagg, R. M. Braziel, D. A. Arber, K. E. Bijwaard, and A. Y. Chu Immunoglobulin Heavy Chain Gene Analysis in Lymphomas: A Multi-Center Study Demonstrating the Heterogeneity of Performance of Polymerase Chain Reaction Assays J. Mol. Diagn., May 1, 2002; 4(2): 81 - 89. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
