help button home button Endocrine Society JCEM
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Submit a related Letter to the Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Takano, T.
Right arrow Articles by Amino, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Takano, T.
Right arrow Articles by Amino, N.
The Journal of Clinical Endocrinology & Metabolism Vol. 85, No. 2 671-675
Copyright © 2000 by The Endocrine Society

Original Studies

Diagnosis of Thyroid Malignant Lymphoma by Reverse Transcription-Polymerase Chain Reaction Detecting the Monoclonality of Immunoglobulin Heavy Chain Messenger Ribonucleic Acid1

Toru Takano, Akira Miyauchi, Fumio Matsuzuka, Hiroshi Yoshida, Kanji Kuma and Nobuyuki Amino

Department of Laboratory Medicine (T.T., N.A.), Osaka University Medical School, D2, 2–2 Yamadaoka, Suita, Osaka 565-0871; and Kuma Hospital (A.M., F.M., H.Y., K.K.), 8–2-35 Simoyamate-Dori, Chuo-Ku, Kobe, Hyogo 650-0011, Japan

Address all correspondence and requests for reprints to: Dr. Toru Takano, Department of Laboratory Medicine, Osaka University Medical School, D2, 2–2, Yamadaoka, Suita, Osaka 565-0871, Japan. E-mail: ttakano{at}labo.med.osaka-u.ac.jp


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Distinguishing between thyroid malignant lymphoma and lymphocytic thyroiditis (Hashimoto’s thyroiditis) is quite difficult and problematic. Molecular techniques to detect clonal lymphoid proliferation based on Ig heavy chain (IgH) gene rearrangement may be used to facilitate more accurate diagnosis of malignant lymphoma. We recently established a method for diagnosing thyroid tumors by analyzing ribonucleic acids (RNAs) extracted from the needles used for fine needle aspiration biopsy (aspiration biopsy-RT-PCR). By applying the aspiration biopsy-RT-PCR method to detection of the monoclonality of IgH messenger RNA (mRNA), an accurate molecular-based diagnosis of malignant lymphoma can be established as an adjunct to cytological diagnosis. We first studied RNAs from fresh tissues samples of 8 cases of Hashimoto’s thyroiditis and 18 malignant lymphomas to detect the monoclonality of IgH mRNA by seminested RT-PCR. Monoclonality was detected in 8 of 18 (44.4%) malignant lymphomas, but in none of the 8 cases of Hashimoto’s thyroiditis. We then studied aspirates from 10 cases of thyroid malignant lymphoma, 4 cases of Hashimoto’s thyroiditis, and 1 case each of adenomatous goiter and papillary carcinoma. Monoclonality was detected in the aspirates from 4 of 10 malignant lymphomas (40%), but not from other tissues. Thus, RT-PCR detection of monoclonality of IgH mRNA in addition to cytological examination may be useful in diagnosing thyroid malignant lymphoma.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
PRIMARY THYROID lymphoma (non-Hodgkin’s lymphoma) is a rare type of malignant tumor accounting for 2.2–5% of all thyroid cancers and typically occurs in elderly women with a history of lymphocytic thyroiditis (Hashimoto’s thyroiditis) (1, 2, 3, 4, 5, 6). This tumor originates from B lymphocytes and rarely from T cells (7). The cytological diagnosis of primary malignant lymphoma of the thyroid, particularly high grade lymphoma, is possible by fine needle aspiration biopsy (FNAB) (8, 9); however, the cytological differentiation of Hashimoto’s thyroiditis from low grade lymphoma can be problematic due to the lack of marked nuclear atypia. The use of immunotyping with B cell-specific monoclonal antibody (10) or flow cytometry (11) may help to identify clonal populations of lymphoid cells indicative of malignant lymphoma, but accurate, efficient, and less invasive methods of diagnosing thyroid lymphomas other than open biopsy have not been firmly established.

Recently, PCR had become a viable alternative to the traditional, but more costly and time-consuming, Southern blot hybridization (SBH) analysis for the assessment of Ig heavy chain gene (IgH) rearrangement (B cell monoclonality) (12, 13, 14). B cell populations from many B cell neoplasms can be readily amplified with a standard approach using a consensus IgH variable region (VH) framework (FR) III-directed primer, along with a consensus IgH joining region (JH) primer. The assays have been shown to be very specific, although the overall monoclonality detection rate, or diagnostic sensitivity, is consistently lower than that of Southern hybridization analysis. However, this method cannot be applied to samples obtained by FNAB, because it requires 1 µg purified genomic DNA for a successful analysis, which cannot be obtainable by the usual FNAB procedures.

When using only a small number of cells, the analysis of messenger ribonucleic acid (mRNA) is much easier than that of DNA because the copy number of each mRNA is usually larger than that of the corresponding DNA. To establish a method of preoperative molecular-based diagnosis of thyroid cancers, we previously introduced a new technique, aspiration biopsy-RT-PCR (ABRP) (15). In this technique, RNAs extracted from tumor cells in the needle used for FNAB are used for RT-PCR. ABRP provides additional RNA analysis data to augment the results of cytological diagnosis. The RNA extracted from an FNAB provides sufficient complementary DNA (cDNA) for as many as 20 PCR examinations. We have demonstrated the clinical usefulness of preoperative molecular-based diagnosis by the ABRP detecting oncofetal fibronectin mRNA, the expression of which is restricted in thyroid papillary and anaplastic carcinomas (16, 17, 18). Further, we also demonstrated that thyroid medullary carcinomas can be successfully preoperatively diagnosed by the ABRP detecting RET protooncogene, calcitonin, and carcinoembryonic antigen (CEA) mRNAs (19). In this study, to establish a simpler and more rapid diagnosing method, we applied ABRP to the preoperative diagnosis of malignant lymphoma by amplifying IgH cDNA to detect its monoclonality and then attempted to clarify its clinical usefulness.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Extraction of RNAs from the tissue samples and FNABs

Twenty-six thyroid tissue samples (18 malignant lymphomas and 8 cases of Hashimoto’s thyroiditis) were obtained by open biopsy (Table 1Go). All malignant lymphomas were B cell lymphomas. A portion of the tissue sample was immediately frozen in liquid nitrogen. Total RNA was extracted according to the method of Chomczynski and Sacchi (20) for use in the following study. FNAB samples from 16 thyroid tissues (10 malignant lymphomas, 4 cases of Hashimoto’s thyroiditis, and 1 case each of papillary carcinoma and adenomatous goiter) were obtained preoperatively by ABRP as previously described (15) (Table 2Go). In brief, a syringe with a 22-gauge needle was used to obtain a FNAB from the tissue sample. A sample of the FNAB was prepared on a slide glass for cytological examination, and leftover cells inside the needle were then lysed with a denaturing solution containing 4 mol/L guanidine thiocyanate, 25 mmol/L sodium citrate (pH 7.0), 0.5% sarcosyl, and 0.1 mol/L 2-mercaptoethanol into a 1.5-mL tube. The tubes were then stored at 4 C. Total cellular RNA was extracted as previously described. Whenever possible, the RNA extracted from the whole aspirate was similarly obtained. After surgery or open biopsy, the diagnoses were confirmed histologically. All malignant lymphomas were B cell lymphomas.


View this table:
[in this window]
[in a new window]
  		Table 1. Malignant lymphomas used in the RT-PCR analysis

 

View this table:
[in this window]
[in a new window]
  		Table 2. Malignant lymphomas used in the ABRP analysis

 
Seminested RT-PCR analysis

RT was performed using either the whole RNA extracted by ABRP or 1 µg total RNA from tissue samples in a RT mixture containing 50 mmol/L Tris-HCl (pH 8.3), 75 mmol/L KCl, 10 mmol/L dithiothreitol, 3 mmol/L MgCl2, 0.5 mmol/L deoxynucleotide triphosphates (dNTPs), 200 U Moloney murine leukemia virus reverse transcriptase (Life Technologies, Inc., Gaithersburg, MD), 2 U/µL ribonuclease inhibitor (Takara, Shiga, Japan), and 2.5 µmol/L random hexamer (Takara) in a total volume of 20 µL at 42 C for 60 min. One microliter of first strand cDNA was used as a template for the seminested PCR reaction with specific primers for IgH cDNA (Fig. 1Go). The sequences of the primers are given in Table 3Go (21). All primers were purchased from Life Technologies, Inc. For the first PCR, each reaction mixture consisted of 1 µL cDNA, 0.5 µmol/L FR3A and LJH, 2 mmol/L MgCl2, 0.3 µL dimethylsulfoxide, 1 µL 10 x PCR buffer II, 1 µL 200 µmol/L dNTP mix, 0.5 U AmpliTaq Gold DNA polymerase, and nuclease-free water to a final volume of 10 µL. 10 x PCR buffer II, dNTP mix, and AmpliTaq Gold DNA polymerase were obtained from Perkin-Elmer Corp./Cetus (Emeryville, CA). The reaction mixture was subjected to the PCR reaction. The conditions were 95 C for 10 min followed by 20 cycles of denaturation (95 C, 1 min), annealing (55 C, 1 min), then final extension at 72 C for 10 min. For the second PCR, each reaction mixture consisted of 0.3 µL of the first PCR sample, 0.5 µmol/L FR3A and VLJH, 2 mmol/L MgCl2, 0.6 µL dimethylsulfoxide, 2 µL 10 x PCR buffer II, 2 µL 200 µmol/L dNTP mix, 1 U AmpliTaq Gold DNA polymerase, and nuclease-free water to a final volume of 20 µL. The PCR conditions were 95 C for 10 min followed by 15 cycles of denaturation (95 C, 1 min), annealing (60 C, 1 min), then final extension at 72 C for 10 min. The PCR products were separated by 12% PAGE. The gel was stained with SYBR Green I (Takara), and the fluorescence image was analyzed with a Fluor Imager (Molecular Dynamics, Inc., Sunnyvale, CA). The sample, which showed a clearly dominant band by seminested RT-PCR, was determined to be positive for monoclonality. In the case of FNAB, the patients who showed monoclonality in either the leftover cells or the whole aspirate were categorized as positive. One microgram of the RNA extracted from human peripheral blood by Isogen-LS (Wako, Osaka, Japan) was used as a negative control, and 1 µg RNA extracted from a B cell lymphoma, which produces a 102-bp band derived from IgH cDNA by RT-PCR, was used as a positive control.



View larger version (7K):
[in this window]
[in a new window]
  		Figure 1. Primer design.

 

View this table:
[in this window]
[in a new window]
  		Table 3. Sequences of oligonucleotide primers

 
SBH

Southern transfer and prehybridization were performed using a digoxigenin DNA labeling kit and a digoxigenin luminescent detection kit for nucleic acids (Roche Molecular Biochemicals, Tokyo, Japan) according to standard protocols that have been previously described (22). In brief, 10 µg DNA extracted from a tissue sample were digested with BamHI and HindIII. The digests were electrophoresed on an 0.8% agarose gel, transferred to a nylon membrane (Hybond-N+, Amersham Pharmacia Biotech, Tokyo, Japan), and hybridized overnight with a 5.4-kb JH genomic DNA probe (Oncogene Science, Inc., Uniondale, NY) labeled with digoxigenin. After probe hybridization, the membrane was incubated in a blocking solution for 2 h at room temperature. The blocking solution was discarded, and antidigoxigenin antibody conjugated to alkaline phosphatase was added. After appropriate washing, the signal was detected by adding disodium-3-(4-methoxyspirol 1,2-dioxethane-3,2'-(5'-chloro)tricyclo[3.3.1.1(3, 7)]decan-4yl)phenyl phosphate (Roche Molecular Biochemicals), and the membrane was then exposed to a radiographic film.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
To evaluate the efficiency of the seminested RT-PCR detection of the monoclonality of IgH mRNA, we amplified IgH cDNA by seminested RT-PCR using the tissue samples. Clear monoclonal bands were observed in 8 of 18 (44.4%) malignant lymphomas, but not in the 8 cases of Hashimoto’s thyroiditis (Fig. 2Go and Table 1Go). Fifteen malignant lymphomas were subjected to SBH analysis. Monoclonality of the IgH gene was observed in 14 of 16 (87.5%) malignant lymphomas. Next, RNAs obtained by FNAB were amplified by seminested RT-PCR to detect the monoclonality of IgH mRNA (Fig. 3Go and Table 2Go). Monoclonality was positive in 4 of 10 (40%) malignant lymphomas, but was negative in all 4 cases of Hashimoto’s thyroiditis as well as the cases of adenomatous goiter and papillary carcinomas. The positive and negative predictive values of this assay were both 100%. In 1 patient, monoclonality was detected only when the whole aspirates were used for the assay. SBH analysis showed monoclonality of the IgH gene in 7 of 9 (77.7%) malignant lymphomas.



View larger version (43K):
[in this window]
[in a new window]
  		Figure 2. Representative results of RT-PCR detection of IgH mRNA monoclonality. Total RNAs extracted from tissue samples were subjected to seminested RT-PCR analysis. The PCR products were separated by 12% PAGE, and the fluorescence image was analyzed with a Fluor Imager. The samples showing a clear dominant band (lanes 6, 7, 8, and 10) were determined to be positive for monoclonality. N, RNA from peripheral blood as a negative control; P, RNA from a B cell malignant lymphoma as a positive control.

 


View larger version (38K):
[in this window]
[in a new window]
  		Figure 3. Representative results of ABRP detection of IgH mRNA monoclonality. Total RNAs extracted from FNABs were subjected to seminested RT-PCR analysis. The PCR products were separated by 12% PAGE. The samples that showing a clear dominant band (lanes 6, 8, and 10) were determined to be positive for monoclonality. N, RNA from peripheral blood as a negative control; P, RNA from a B cell malignant lymphoma as a positive control.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Most patients with malignant lymphoma note a rapidly growing mass in the neck over a period of 2–3 months, and symptoms of thyroid lymphoma are mainly due to rapid growth of the tumor with compression of the adjacent organs (23). However, these symptoms often appear in patients with the aggressive inflammation caused by Hashimoto’s thyroiditis, and Hashimoto’s thyroiditis is believed to be one of the causes of malignant lymphoma. Thus, clinical differentiation between these two conditions is quite difficult, and in most cases a histopathological method by open biopsy is used to confirm the diagnosis. Molecular-based preoperative diagnosis is expected to reduce the invasion to the patients, although there have been only a few studies based on PCR analysis and no studies on RT-PCR analysis of thyroid malignant lymphoma. Lovchik et al. reported a case of malignant lymphoma in which a clonal population of lymphocytes within the FNA specimen was identified using a PCR-based assay for IgH gene rearrangement (24), but no other report of molecular-based diagnosis of malignant lymphoma using FNABs has been published. This is partly because a malignant lymphoma in the thyroid is a rare tumor, and the fresh tissue or FNAB samples that can be used for RNA analysis are seldom obtainable.

In this study we collected RNAs from 18 tissue samples and 10 FNABs obtained from tumors histopathologically diagnosed as malignant lymphoma to estimate the clinical usefulness of detection of the monoclonality of IgH mRNA. In studies of other groups, PCR-based B cell clonality analyses using DNA as the template show a sensitivity and specificity of about 60–80% and 100%, respectively (25, 26, 27, 28). In this study using seminested RT-PCR, the specificity was 100%, but the sensitivity using the tissue samples was 44.4%, which is considerably lower than that obtained when using DNA, although the specificity should be more extensively examined in an additional study because only 8 cases of Hashimoto’s thyroiditis have been tested. The low expression level of IgH mRNA in malignant lymphoma can be a cause of low sensitivity. However, this does not seem to be likely, because previous studies using immunohistochemistry have detected the expression of Ig in the majority of B cell lymphomas (29, 30). Before this study, we attempted to detect the monoclonality of IgH cDNA by single RT-PCR. This trial failed, because it often produced many false positive bands that were not derived from IgH cDNAs (data not shown). Thus, we found that when using RNAs, we should choose more restricted RT-PCR amplification conditions to detect a true monoclonal band. Recently, using RT-PCR-single strand conformation polymorphism, Shiokawa et al. reported a similar sensitivity (50%) in detecting the monoclonality of IgH mRNA in 16 cases of B cell lymphoma (31). Considering these previously reported results, the low sensitivities in the present study may be a consequence of significant mismatch (i.e. poor sequence homology) between VH primer (FR3A) and the VH-FR III region of a particular B cell lymphoma or a deletion of the terminal 3'-bases of this VH-FR III region (32, 33), because PCR was carried out in more restricted amplification conditions than those in the previous studies. If a modification of this method, in which IgH cDNA is amplified without using a primer annealing to the VH sequence, is possible, it may improve the sensitivity.

In ABRP detection of the monoclonality of IgH mRNA, we have not experienced any false positive results, although only six nonlymphomatous thyroid aspirates have been studied. However, the sensitivity of this method in malignant lymphoma appeared to be almost the same as that using tissue samples (40%). In one case, monoclonality was only detected when the whole aspirate was used for the analysis. In FNAB, much contamination of peripheral blood cells occurs frequently, and the considerable volume of contaminated blood cells may prevent detection of the monoclonality of IgH mRNA. Further, some cases showed discrepant results between ABRP and SBH. This may be at least in a part because the part of the tissue used for SBH was not always the same as that used for aspiration. Thus, in the case of molecular-based diagnosis of malignant lymphoma, multiple examinations by FNAB, as recommended by Hamburger et al. (34), may be expected to prevent false negative results.

By using combined analyses of molecular and cytological approaches, about half of malignant lymphomas may be diagnosed by FNAB without using more invasive methods, such as open biopsy. Further, the rapid detection of monoclonality of IgH mRNA by RT-PCR, which takes only a half a day, may help cytopathologists to decide whether the patient needs further examination to confirm the diagnosis of malignant lymphoma.


   Footnotes
 
1 This work was supported by a Grant-in-Aid for Encouragement of Young Scientists (to T.T.; no. 10771346) from the Ministry of Education, Science, Sports, and Culture of Japan and a Grant-in-Aid from Kurozumi Medical Foundation. Back

Received May 24, 1999.

Revised October 22, 1999.

Accepted November 3, 1999.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Hamburger JI, Miller JM, Kini SR. 1983 Lymphoma of the thyroid. Ann Intern Med. 99:685–693.
  2. Rasbach DA, Mondschein MS, Harris NL, Kaufman DS, Wang CA. 1985 Malignant lymphoma of the thyroid gland: a clinical and pathologic study of twenty cases. Surgery. 98:1166–1170.[Medline]
  3. Anscombe AM, Wright DH. 1985 Primary malignant lymphoma of the thyroid–a tumour of mucosa- associated lymphoid tissue: review of seventy-six cases. Histopathology. 9:81–97.[Medline]
  4. Kato I, Tajima K, Suchi T, et al. 1985 Chronic thyroiditis as a risk factor of B-cell lymphoma in the thyroid gland. Jpn J Cancer Res. 76:1085–1090.[Medline]
  5. Tupchong L, Hughes F, Harmer CL. 1986 Primary lymphoma of the thyroid: clinical features, prognostic factors, and results of treatment. Int J Radiat Oncol Biol Physiol. 12:1813–1821.[Medline]
  6. Aozasa K. 1990 Hashimoto’s thyroiditis as a risk factor of thyroid lymphoma. Acta Pathol Jpn. 40:459–468.[Medline]
  7. Aozasa K, Ueda T, Katagiri S, Matsuzuka F, Kuma K, Yonezawa T. 1987 Immunologic and immunohistologic analysis of 27 cases with thyroid lymphomas. Cancer. 60:969–973.[CrossRef][Medline]
  8. Willems JS, Lowhagen T. 1981 The role of fine-needle aspiration cytology in the management of thyroid disease. Clin Endocrinol Metab. 10:267–273.[CrossRef][Medline]
  9. Matsuda M, Sone H, Koyama H, Ishiguro S. 1987 Fine-needle aspiration cytology of malignant lymphoma of the thyroid. Diagn Cytopathol. 3:244–249.[Medline]
  10. Samaan NA, Ordonez NG. 1990 Uncommon types of thyroid cancer. Endocrinol Metab Clin North Am. 19:637–648.[Medline]
  11. Zirker TA, Baybick JH, Vincent Jr JL, Smith GJ, Siegal GP. 1988 Flow cytometric analysis of DNA ploidy in lymphomas of the thyroid. Head Neck Surg. 10:324–329.[Medline]
  12. McCarthy KP, Sloane JP, Wiedemann LM. 1990 Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens. J Clin Pathol. 43:429–432.[Abstract/Free Full Text]
  13. Trainor KJ, Brisco MJ, Story CJ, Morley AA. 1990 Monoclonality in B-lymphoproliferative disorders detected at the DNA level. Blood. 75:2220–2222.[Abstract/Free Full Text]
  14. Deane M, Norton JD. 1990 Detection of immunoglobulin gene rearrangement in B lymphoid malignancies by polymerase chain reaction gene amplification. Br J Haematol. 74:251–256.[Medline]
  15. Takano T, Sumizaki H, Amino N. 1997 Detection of CD44 variants in fine needle aspiration biopsies of thyroid tumor by RT-PCR. J Exp Clin Cancer Res. 16:267–271.[Medline]
  16. Takano T, Matsuzuka F, Sumizaki H, Kuma K, Amino N. 1997 Rapid detection of specific messenger RNAs in thyroid carcinomas by reverse transcription-PCR with degenerate primers: specific expression of oncofetal fibronectin messenger RNA in papillary carcinoma. Cancer Res. 57:3792–3797.[Abstract/Free Full Text]
  17. Takano T, Matsuzuka F, Miyauchi A, et al. 1998 Restricted expression of oncofetal fibronectin mRNA in thyroid papillary and anaplastic carcinoma: an in situ hybridization study. Br J Cancer. 78:221–224.[Medline]
  18. Takano T, Miyauchi A, Yokozawa T, et al. 1998 Accurate and objective preoperative diagnosis of thyroid papillary carcinomas by reverse transcription-PCR detection of oncofetal fibronectin messenger RNA in fine-needle aspiration biopsies. Cancer Res. 58:4913–4917.[Abstract/Free Full Text]
  19. Takano T, Miyauchi A, Matsuzuka F, et al. 1999 Preoperative diagnosis of medullary thyroid carcinoma by RT-PCR using RNA extracted from leftover cells within a needle used for fine needle aspiration biopsy. J Clin Endocrinol Metab. 84:951–955.[Abstract/Free Full Text]
  20. Chomczynski P, Sacchi N. 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate- phenol-chloroform extraction. Anal Biochem. 162:156–159.[Medline]
  21. Ramasamy I, Brisco M, Morley A. 1992 Improved PCR method for detecting monoclonal immunoglobulin heavy chain rearrangement in B cell neoplasms. J Clin Pathol. 45:770–775.[Abstract/Free Full Text]
  22. Cleary ML, Chao J, Warnke R, Sklar J. 1984 Immunoglobulin gene rearrangement as a diagnostic criterion of B-cell lymphoma. Proc Natl Acad Sci USA. 81:593–597.[Abstract/Free Full Text]
  23. Matsuzuka F, Miyauchi A, Katayama S, et al. 1993 Clinical aspects of primary thyroid lymphoma: diagnosis and treatment based on our experience of 119 cases. Thyroid. 3:93–99.[Medline]
  24. Lovchik J, Lane MA, Clark DP. 1997 Polymerase chain reaction-based detection of B-cell clonality in the fine needle aspiration biopsy of a thyroid mucosa-associated lymphoid tissue (MALT) lymphoma. Hum Pathol. 28:989–992.[CrossRef][Medline]
  25. Diss TC, Peng H, Wotherspoon AC, Isaacson PG, Pan L. 1993 Detection of monoclonality in low-grade B-cell lymphomas using the polymerase chain reaction is dependent on primer selection and lymphoma type. J Pathol. 169:291–295.[CrossRef][Medline]
  26. Slack DN, McCarthy KP, Wiedemann LM, Sloane JP. 1993 Evaluation of sensitivity, specificity, and reproducibility of an optimized method for detecting clonal rearrangements of immunoglobulin and T-cell receptor genes in formalin-fixed, paraffin-embedded sections. Diagn Mol Pathol. 2:223–232.[Medline]
  27. Reed TJ, Reid A, Wallberg K, O’Leary T., Frizzera G. 1993 Determination of B-cell clonality in paraffin-embedded lymph nodes using the polymerase chain reaction. Diagn Mol Pathol. 2:42–49.[Medline]
  28. Ilyas M, Jalal H, Linton C, Rooney N. 1995 The use of the polymerase chain reaction in the diagnosis of B-cell lymphomas from formalin-fixed paraffin-embedded tissue. Histopathology. 26:333–338.[Medline]
  29. Warnke R, Miller R, Grogan T, Pederson M, Dilley J, Levy R. 1980 Immunologic phenotype in 30 patients with diffuse large-cell lymphoma. N Engl J Med. 303:293–300.[Abstract]
  30. Strauchen JA, Mandeli JP. 1991 Immunoglobulin expression in B-cell lymphoma. Immunohistochemical study of 345 cases. Am J Clin Pathol. 95:692–695.[Medline]
  31. Shiokawa S, Nishimura J, Ohshima K, Uike N, Yamamoto K. 1998 Establishment of a novel B cell clonality analysis using single-strand conformation polymorphism of immunoglobulin light chain messenger signals. Am J Pathol. 153:1393–1400.[Abstract/Free Full Text]
  32. Cleary ML, Meeker TC, Levy S, et al. 1986 Clustering of extensive somatic mutations in the variable region of an immunoglobulin heavy chain gene from a human B cell lymphoma. Cell. 44:97–106.[CrossRef][Medline]
  33. Deane M, McCarthy KP, Wiedemann LM, Norton JD. 1991 An improved method for detection of B-lymphoid clonality by polymerase chain reaction. Leukemia. 5:726–730.[Medline]
  34. Hamburger JI, Husain M, Nishiyama R, Nunez C, Solomon D. 1989 Increasing the accuracy of fine-needle biopsy for thyroid nodules. Arch Pathol Lab Med. 113:1035–1041.[Medline]



This article has been cited by other articles:


Home page
 J. Clin. Endocrinol. Metab.Home page
T. Takano, A. Miyauchi, F. Matsuzuka, H. Yoshida, T. Notomi, K. Kuma, and N. Amino
Detection of Monoclonality of the Immunoglobulin Heavy Chain Gene in Thyroid Malignant Lymphoma by Vectorette Polymerase Chain Reaction
J. Clin. Endocrinol. Metab., February 1, 2005; 90(2): 720 - 723.
[Abstract] [Full Text] [PDF]

Home page
 Jpn J Clin OncolHome page
T. Takano, Y. Hasegawa, A. Miyauchi, F. Matsuzuka, H. Yoshida, K. Kuma, and N. Amino
Quantitative Analysis of Thymosin Beta-10 Messenger RNA in Thyroid Carcinomas
Jpn. J. Clin. Oncol., July 1, 2002; 32(7): 229 - 232.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
C. Thieblemont, A. Mayer, C. Dumontet, Y. Barbier, E. Callet-Bauchu, P. Felman, F. Berger, X. Ducottet, C. Martin, G. Salles, et al.
Primary Thyroid Lymphoma Is a Heterogeneous Disease
J. Clin. Endocrinol. Metab., January 1, 2002; 87(1): 105 - 111.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Submit a related Letter to the Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Takano, T.
Right arrow Articles by Amino, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Takano, T.
Right arrow Articles by Amino, N.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals