IgH和IgL引物联合检测对提高石蜡组织淋巴瘤基因重排检出率的价值

背景与目的：通过聚合酶链反应（polymerase chain reaction,PCR）扩增免疫球蛋白重链（immunoglobulin heavy chain,IgH）基因对其克隆性的检测,可以辅助诊断淋巴瘤。缺点是假阴性率较高,在石蜡包埋组织中尤为明显。本研究拟采用手工显微切割、免疫球蛋白重链和轻链（immunoglohulin light chain,IgL）联合测定等方式,探讨该方法在石蜡包埋组织中非霍奇金淋巴瘤（non-Hodgkin’s lymphoma,NHL）诊断中的价值。方法：选用1对IgH引物、1对T细胞受体γ（Tcell receptor γ,TCRγ）、TCRγ引物、2对新设计的轻链引物,通过PCR、琼脂糖和聚丙烯酰胺凝胶电泳（PAGE）及银染技术,检测经形态学及免疫组织化学确诊的58例石蜡包埋组织标本,包括39例B细胞淋巴瘤、16例T细胞淋巴瘤和3例淋巴结反应性增生组织。以DG75和Jurkat淋巴瘤细胞系DNA作为对照。结果：IgH引物P1在39例B细胞淋巴瘤检出阳性率79．5％（31／39）。假阳性率6．25％（1／16）,IgL引物在B细胞淋巴瘤检出阳性率71．8％（28／39）。假阳性率12．5％（2／16）,经统计学分析二者检出率无显著性差异（P〉0．05）。二者联合检测,B细胞淋巴瘤阳性检出率可以达到92．3％,假阳性率并无明显提高（12．5％）。以上重排引物在反应性增生淋巴结组织中均未检出。结论：IgH与IgL引物联合检测可明显提高石蜡包埋组织中B细胞淋巴瘤的检出率,并为B-NHL的诊断及鉴别诊断提供了有效的辅助手段。     

B细胞淋巴瘤患者骨髓IgH基因克隆性重排检测的临床意义

目的 探讨半巢式聚合酶链反应(PCR)检测B细胞淋巴瘤患者骨髓中IgH基因克隆性重排的可行性,并初步评价其临床价值.方法 选用FR2、FR3A引物,采用半巢式PCR方法检测105例B细胞淋巴瘤患者骨髓中IgH基因的单克隆性重排,与骨髓穿刺细胞形态学检测结果进行比较,并评价PCR检测结果与临床病理特征的关系.结果 105例B细胞淋巴瘤患者中,IgH基因克隆性重排PCR检测48例(45.7%)阳性,而骨髓细胞形态学只检测出22例(21.0%),两者差异有统计学意义(P<0.05),符合率为71.4%(75/105).弥漫大B细胞性淋巴瘤(DLBCL)、滤泡性淋巴瘤(FL)及小淋巴细胞性淋巴瘤(SLL)初治患者PCR检测阳性率分别为30.8%、25.0%和100.0%.PCR检测结果与Ann Arbor分期有关,早期B细胞淋巴瘤患者lgH基因克隆性重排PCR检出阳性率低于晚期患者(P=0.02).PCR检测阳性和阴性患者的近期疗效差异无统计学意义(P>0.05),但CR率(23.3%和46.3%)差异有统计学意义(P=0.019).结论 IgH基因克隆性重排PCR检测可能是判断B细胞淋巴瘤患者骨髓异常的有效方法,较骨髓细胞形态学敏感;Ann Arbor分期晚的患者PCR检测阳性率高于分期早的患者;PCR检测阳性者治疗后获得CR的机会低于阴性者.     

聚合酶链反应检测非霍奇金淋巴瘤克隆性基因重排的临床应用

目的建立聚合酶链反应（ＰＣＲ）克隆基因重排技术，探讨其对非霍奇金淋巴瘤（ＮＨＬ）疑难病例的诊断价值。方法对疑难ＮＨＬ病例标本２１例和儿科淋巴细胞白血病治疗后残留病变标本１１例进行免疫球蛋白重链（ＩｇＨ）和Ｔ细胞受体（ＴＣＲβ）克隆性基因重排检测。结果２１例疑难病例全部阳性（１０％），１１例淋巴细胞白血病治疗后残留病变中有９例（８２％）阳性。结论ＰＣＲ技术可用于临床病理活检ＮＨＬ疑难病例的辅助诊断。     

The detection of specific gene rearrangements in non-Hodgkin's lymphoma using the polymerase chain reaction.

Characteristic gene rearrangements are present in most non-Hodgkin's lymphomas (NHL). These are usually detected by Southern blotting techniques. In this study, the ability of the polymerase chain reaction (PCR) to detect the t(14;18) chromosomal translocation and immunoglobulin heavy chain (IgH) gene rearrangement was evaluated. DNA from 14 follicular and 42 diffuse B-cell lymphomas was examined using oligonucleotide primers specific for opposing sides of the IgH gene rearrangement on chromosome 14 (towards conserved VH and JH sequences) and opposing sides of the t(14;18) chromosomal translocation (towards the major breakpoint region of the bcl-2 gene on chromosome 18 and conserved JH sequence on chromosome 14). The t(14;18) translocation was detected in 57% of follicular lymphomas and 21% of diffuse B-cell lymphomas. Clonal IgH gene rearrangements using PCR were detected in 50% follicular and 52% of the diffuse lymphomas. Either or both of these rearrangements were detected in 93% follicular and in 59% of diffuse lymphomas. PCR is a rapid and easy technique that can detect the abnormal rearrangement of the bcl-2 gene and clonal IgH rearrangement, indicating the presence of lymphoma. This may be of benefit in monitoring response to therapy and in predicting prognosis in this disease.     

Application of laser capture microdissection to cytologic specimens for the detection of immunoglobulin heavy chain gene rearrangement in patients with malignant lymphoma

BACKGROUND: The demonstration of the monoclonality of immunoglobulin heavy chain (IgH) gene rearrangement is an indispensable method for the diagnosis of B-cell lymphoma as well as histocytochemical analysis. For the detection of IgH gene rearrangement, the extraction of DNA from a homogenous cell population is necessary. Recently, the laser capture microdissection (LCM) technique was shown to isolate specific cells from histopathologic specimens for molecular analysis. However, to the authors' knowledge the applicability of LCM to cytologic specimens has not yet been well established. METHODS: Using LCM, a homogenous population of B-cell lymphoma cells as both histologic sections and cytologic specimens was captured, and genomic DNA was extracted from the captured cells. IgH gene rearrangement was analyzed by the polymerase chain reaction (PCR)-based single-strand conformational polymorphism (SSCP) method. RESULTS: Genomic DNAs were extracted successfully from ethanol-fixed cytologic specimens, but cells were not captured from air-dried specimens. Using PCR-SSCP analysis, the monoclonality of the IgH gene rearrangement was detected in five cases of tissue sections among nine analyzed cases of malignant lymphoma diagnosed immunohistochemically. However, analysis of the cytologic specimens with LCM demonstrated the monoclonality of the IgH gene rearrangement in seven cases of lymphoma. CONCLUSIONS: The results of the current study suggest that the novel application of LCM to cytologic specimens occasionally exhibits high sensitivity for the detection of IgH gene rearrangement monoclonality compared with the use of histologic sections.     

Detection of clonality in lymphoproliferations using PCR of the antigen receptor genes: does size matter?

A biopsy of a nasal mass that had morphologic and immunostaining features consistent with a B-cell lymphoma was studied for clonality using PCR of the IgH gene. An unexpectedly low molecular weight DNA fragment of approximately 140bp (acceptable size limit: 250-295bp) was obtained using FR2 and JH primers. The sequence of this DNA was consistent with a clonal IgH rearrangement followed by a deletion that removed most of the downstream portion of the V segment. Thus, the biopsy contained a monoclonal population of B-lymphocytes, consistent with a diagnosis of lymphoma. This work illustrates that bands outside of the size range expected from PCR of the antigen receptor genes may still be consistent with a monoclonal result.     

结节性淋巴细胞为主型霍奇金淋巴瘤和富于T细胞和(或)组织细胞的B细胞淋巴瘤的鉴别诊断

目的 探讨结节性淋巴细胞为主型霍奇金淋巴瘤（NLPHL）和富于T细胞和（或）组织细胞的B细胞淋巴瘤（TCRBCL）的鉴别诊断。方法 按照WHO淋巴瘤新分类法,对15例NLPHL和16例TCRBCL的组织学和免疫表型进行分析,并分别对其中3例NLPHL和4例TCRBCL进行了EBERl／2原位杂交和IgH基因重排检测。结果 组织学上,NLPHL和TCRBCL均表现为小淋巴细胞背景中散在分布的肿瘤性大细胞。NLPHL的肿瘤性大细胞形态特征以L＆H细胞（即爆米花细胞）为主,TCRBCL的肿瘤性大细胞以中心母细胞类型为主,两者都可伴有其他变异形态。免疫表型上,NLPHL和TCRBCL的肿瘤性大细胞都呈CD20、CD79a、bcl-6和EMA阳性,CD3、CD45RO、CDl5和CD30阴性,背景小淋巴细胞以T淋巴细胞为主。但在NLPHL中,CD57阳性细胞明显多于TIA-1阳性细胞,小B淋巴细胞呈小灶状或弥漫散在分布;而在TCRBCL中,TIA-1阳性细胞明显多于CD57阳性细胞,小B淋巴细胞非常稀少。CD21检测显示,NLPHL的结节呈CD21阳性滤泡树突细胞（FDC）网架结构,而TCRBCL以及NLPHL的弥漫类型或弥漫区域中,FDC网架缺乏。NLPHL和TCRBCL都呈EBER1／2阴性,IgH基因重排可检测到80～120bp的单克隆条带。结论 NLPHL和TCRBCL有组织学和免疫表型特征的相似性,诊断和鉴别诊断必须结合形态学和瘤细胞、背景细胞的免疫表型特征。     

PCR和FISH技术在子宫颈淋巴瘤与淋巴瘤样病变诊断和鉴别诊断中的作用

 目的　探讨PCR和FISH检测技术在原发性子宫颈淋巴瘤与淋巴瘤样病变的诊断与鉴别诊断中的作用。方法　收集3例原发性子宫颈弥漫性大B细胞淋巴瘤（DLBCL）与2例子宫颈淋巴瘤样病变,进行PCR-IgH重排及FISH检测。结果　PCR检测显示3例DLBCL和1例淋巴瘤样病变中出现单克隆性IgH基因重排。间期FISH检测显示3例DLBCL均发生了IgH和bcl-6基因断裂,而2例淋巴瘤样病变均未检测到特定的染色体断裂。结论　PCR-IgH重排并非仅见于子宫颈细胞淋巴瘤。间期FISH检测IgH和bcl-6基因的断裂对于子宫颈B细胞淋巴瘤和淋巴瘤样病变的诊断和鉴别诊断有帮助。     

Polymerase chain reaction-based diagnosis of bone marrow involvement in 170 cases of non-Hodgkin lymphoma

BACKGROUND: Up to the current time, diagnosis of bone marrow (BM) involvement in non-Hodgkin lymphoma (NHL) has been based on morphologic findings. Polymerase chain reaction (PCR) for antigen receptor gene rearrangements has the potential to increase the detection sensitivity of minimal degrees of BM involvement. The authors therefore assessed PCR-based clonalities of BM concurrently with morphology from 170 cases with NHL and evaluated the usefulness of comparative analysis of clonalities between bilateral BMs and the lymph node and the clinical significance of PCR based clonalities of BM. METHODS: Bilateral BM clot sections of 170 cases and 47 lymph nodes were tested for immunoglobulin heavy chain gene rearrangement or T-cell receptor gamma gene rearrangement according to the B- or T-lineage of the lymph node. RESULTS: When compared with morphology, the results of PCR showed an unexpectedly low positive concordance rate of 61.0% for B-cell NHL and 57.1% for T-cell NHL. When the clonality of BM was compared with that of lymph nodes in B-cell NHL, bilateral clonalities of BM showed high concordance with the clonality of the lymph nodes. PCR-based clonality did not show significant impact on survival. CONCLUSIONS: Morphology remains the gold standard in the evaluation of BM involvement by NHL. Although the comparative analysis of BM clonality and that of the lymph nodes is considered a valuable tool that increases the reliability of clonality, PCR-based clonality of BM does not significantly add to the sensitivity of diagnosing BM involvement by NHL.     

Bronchial infiltration with diffuse large B-cell lymphoma

Non-Hodgkin's lymphoma (NHL) refers to a heterogeneous group of lymphoproliferative diseases with a diversity of clinical courses, including involvement in another organs. NHL frequently involves the thoracic structures, and particularly the mediastinum and lung parenchyma. Several clinical reports have described bronchial-associated lymphoid tissue (BALT) lymphoma as an endobronchial lesion, but endobronchial infiltration with diffuse large B-cell lymphoma is extremely rare. Here, we provide the first report of this condition confirmed by a histopathological study and the presence of an immunoglobulin heavy chain (IgH) gene rearrangement detected by a polymerase chain reaction (PCR).     

Application of Immunophenotyping and Heteroduplex Polymerase Chain Reaction (hPARR) for Diagnosis of Canine Lymphomas

Background: Canine malignant lymphoma is classified into B- or T-cell origin, as in the human case. Due to differences in prognosis, a suitable method needs to be developed for lineage identification. Aims: To determine the accuracy of immunophenotypic and molecular information between three methods: immunocytochemistry (ICC), immunohistochemistry (IHC) and heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) in spontaneous canine lymphomas. Materials and Methods: Peripheral blood, fine needle aspiration and tissue biopsies from enlarged peripheral lymph nodes prior to treatment of 28 multicentric lymphoma patients were collected. Cytopathology and histopathology were examined and classified using the updated Kiel and WHO classifications, respectively. Anti-Pax5 and anti-CD3 antibodies as B- and T-cell markers were applied for immunophenotyping by ICC and IHC. Neoplastic lymphocytes from lymph node and white blood cell pellets from peripheral blood were evaluated by hPARR. Results: In this study, low grade B-cell lymphoma accounted for 25% (7/28), high grade B-cell lymphoma for 64.3% (18/28) and high grade T-cell lymphoma for 10.7% (3/28). According to the WHO classification, 50% of all cases were classified as diffuse large B-cell lymphoma. In addition, ICC showed concordant results with IHC; all B-cell lymphomas showed Pax5+/CD3, and all T-cell lymphomas exhibited Pax5-/CD3+. In contrast to hPARR, 12 B-cell lymphomas featured the IgH gene; seven presented the TCRγ gene; five cases showed both IgH and TCRγ genes, and one case were indeterminate. Three T-cell lymphomas showed the TCRγ gene. The percentage agreement between hPARR and ICC/IHC was 60%. Conclusions: Immunophenotyping should not rely on a single method. ICC or IHC with hPARR should be used concurrently for immunophenotypic diagnosis in canine lymphomas.     

The Use of Immunohistochemistry Improves the Diagnosis of Small Cell Lung Cancer and Its Differential Diagnosis. An International Reproducibility Study in a Demanding Set of Cases

IntroductionThe current WHO classification of lung cancer states that a diagnosis of SCLC can be reliably made on routine histological and cytological grounds but immunohistochemistry (IHC) may be required, particularly (1) in cases in which histologic features are equivocal and (2) in cases in which the pathologist wants to increase confidence in diagnosis. However, reproducibility studies based on hematoxylin and eosin–stained slides alone for SCLC versus large cell neuroendocrine carcinoma (LCNEC) have shown pairwise κ scores ranging from 0.35 to 0.81. This study examines whether judicious use of IHC improves diagnostic reproducibility for SCLC.MethodsNineteen lung pathologists studied interactive digital images of 79 tumors, predominantly neuroendocrine lung tumors. Images of resection and biopsy specimens were used to make diagnoses solely on the basis of morphologic features (level 1), morphologic features along with requested IHC staining results (level 2), and all available IHC staining results (level 3).ResultsFor the 19 pathologists reading all 79 cases, the rate of agreement for level 1 was 64.7%, and it increased to 73.2% and 77.5% in levels 2 and 3, respectively. With IHC, κ scores for four tumor categories (SCLC, LCNEC, carcinoid tumors, and other) increased in resection samples from 0.43 to 0.60 and in biopsy specimens from 0.43 to 0.64.ConclusionsDiagnosis using hematoxylin and eosin staining alone showeds moderate agreement among pathologists in tumors with neuroendocrine morphology, but agreement improved to good in most cases with the judicious use of IHC, especially in the diagnosis of SCLC. An approach for IHC in the differential diagnosis of SCLC is provided.     

Cytologic diagnosis of Burkitt lymphoma

BACKGROUND: The diagnosis and classification of lymphoma require correlation of morphologic, immunophenotypic, and molecular-cytogenetic studies. Fine-needle aspiration biopsy (FNAB) is a valuable diagnostic technique that allows material to be collected for these ancillary studies, and for morphologic evaluation. METHODS: The authors report a series of seven cases clinically or morphologically suspicious for Burkitt lymphoma. Fluorescence in situ hybridization studies (FISH) for c-myc were performed on FNAB material and correlated with cytologic and immunophenotypic data. RESULTS: Six of seven specimens were positive for c-myc rearrangement by FISH. However, only three of these cases represented Burkitt lymphoma, with one additional case of atypical Burkitt lymphoma. The other cases included diffuse large B-cell lymphoma, monomorphic posttransplant B-cell lymphoma, and an aggressive B-cell lymphoma, with the latter case negative for c-myc rearrangement by FISH. Of 2 non-Burkitt lymphoma specimens tested, 1 was positive for the immunoglobulin H/bcl-2 rearrangement, in addition to the c-myc rearrangement, suggesting transformation from a lower grade lymphoma. CONCLUSIONS: These cases illustrated the value of FNAB in the diagnosis of Burkitt lymphoma, as well as the importance of obtaining material for, and integrating results of, ancillary studies for the final diagnosis.     

An advantageous method to evaluate IgH rearrangement and its role in minimal residual disease detection

A sensitive, safe and cheap method to detect minimal residual disease (MRD) is here presented. The PCR-GS technique includes: (a) a fluorescent PCR for the IgH region with CDR3/JH consensus primers; (b) the electrophoresis on an automatic sequencer (ABI PRISM 310); (c) the analysis of results by the GeneScan program. A total of 72 samples were analysed: 34/49 B-cell Non-Hodgkin's Lymphoma (NHL) (69%), six out of seven Multiple Myeloma (MM) (86%), 1/2 Hodgkin's Disease (HD) and 4/4 Acute Lymphoblastic Leukaemia (ALL) were found to be positive, showing a monoclonal IgH rearrangement. The major bias of the PCR-GS method are the 21% of false negatives, but 13/15 negative patients carried t(14;18); consequently, the association of the evaluation by PCR assays of the IgH and BCL2/JH rearrangement allowed to detect a molecular marker of B-neoplasia in more than 94% of tested samples.     

PCR-SSCP检测非霍奇金淋巴瘤患者骨髓克隆性T细胞受体基因重排及其临床意义

背景与目的：进一步了解非霍奇金淋巴瘤（non-Hodgkin‘s lymphoma,NHL)患者骨髓标本克隆T细胞受体（T cell receptor,TCR)基因重排情况及临床意义。方法：应用聚合酶链反应（polymerase chain reaction,PCR)联合单链构象多态性（single-strand conformation polymorphism,SSCP),分析43例NHL患者骨髓标本TCRVγI-Jγ基因重排情况。结果：43例NHL患者有26例（60．5％）存在克隆性TCRVγI－Jγ基因重排。16例骨髓形态学检查未发现淋巴瘤细胞浸润者中有3例（18．8％）发现克隆性TCRVγ－Jγ基因重排。此3例患者分别于4－9个月后行骨髓形态检查时发现淋巴瘤细胞浸润。27例骨髓形态学检查有淋巴瘤细胞浸润者有23例（85．2％）存在克隆性TCRVγI－Jγ基因重排阳性,26例PCR扩增阳性病例经SSCP分析发现7例（26．9）％存在寡／亚克隆重排。7例存在寡／亚克隆重排患者经3－11个月有5例（71．4％）发展为白血病,存在寡／亚克隆重排NHL患者1年内转化为白血病的发生率显著高于无寡／亚克隆重排患者（10．5％）（P＜0．005）。结论：应用PCR检测NHL患者骨髓标本克隆性TCRVγ－Jγ基因重排可较骨髓形态学检查更早发现NHL患者骨髓浸润微小病灶。具有寡／亚克隆NHL患者更易发展为白血病,还可发展为急性非淋巴细胞白血病。     

免疫球蛋白kappa基因重排在B细胞增生性疾病中的检测及其应用

目的：评价免疫球蛋白kappa基因重排检测在B淋巴细胞增生性疾病中的价值。方法：选取93例B淋巴细胞增生性疾病及反应性增生标本(其中包括43例甲醛固定,石蜡包埋标本),运用降落PCR方法扩增Igkappa CDR3区,以异源双链分析鉴别扩增产物以分辨其克隆性。结果：在所有确诊的72例B淋巴细胞增生性疾病中Igkappa扩增的阳性率为47％,其中14例急性B淋巴细胞白血病中有2例阳性(14％);3例慢性淋巴细胞白血病申有2例阳性(67％);55例B细胞恶性淋巴瘤申有30例阳性(55％)。在11例不典型增生的病例中,IgK单克隆检出率为2／11(18％)。10例反应性增生中无一出现单克隆重排。石蜡组织中扩增成功率为32／43(74％)。结论：作为一个独立的分子标志,以PCR方法扩增Igkappa确定B淋巴细胞增生性疾病的克隆性在其临床诊断及生物学特性研究方面是一种有前途的方法,且适用于石蜡标本以进行回顾性研究。     

Monitoring of interobserver agreement in nuclear atypia scoring of node-negative breast carcinomas judged at individual collaborating hospitals in the National Surgical Adjuvant Study of Breast Cancer (NSAS-BC) protocol

BACKGROUND: In the NSAS-BC protocol, the nuclear atypia and mitotic counts are to be judged by pathologists at each participating hospital for assessing high-risk node-negative breast cancers. Therefore, maintenance of interobserver agreement in diagnosis at a higher level is mandatory during the period of patient entry. METHODS: Individual collaborating pathologists originally evaluated the histological eligibility of 107 cases. Three panel pathologists determined consensus diagnoses and 29-37 collaborating pathologists determined modal diagnoses of these cases at three slide conference sessions. The original diagnoses were compared with the consensus and modal diagnoses to estimate the percentage of erroneous judgments. RESULTS: The agreement rate in histological type and nuclear atypia score was 69% (74/107) between the original and consensus diagnoses, 76% (81/107) between the original and modal diagnoses and 86% (92/107) between the consensus and modal diagnoses. The strength of interobserver agreement at the slide conference sessions was moderate (0.447-0.535) by kappa statistics. The original, consensus and modal diagnoses were concordant in 71 cases (66%), but were discordant in 36. Of 35 invasive ductal carcinomas with discordant diagnoses, the discordance arose from the intermediate tumor nature in 15, multiple factors in 13 and erroneous diagnosis in seven (6.5%), if the characteristics of the tumor were judged from the percentage interobserver agreement per tumor at the slide conferences. CONCLUSION: Nuclear atypia scoring given at individual hospitals on case entry was almost reproducible among the pathologists. Continuous efforts are needed to improve interobserver agreement and to decrease erroneous diagnosis for protocol eligibility.     

Estrogen and progesterone receptor assay in paraffin-embedded breast cancer--reproducibility of assessment

Immunohistochemical (IHC) assays are routinely used for determining the estrogen receptor (ER) and progesterone receptor (PgR) status of women with breast cancer. The present knowledge of reproducibility of the subjective IHC assessment is limited. The purpose of this study was to evaluate the interobserver reproducibility in IHC assessment of 127 (ER) and 126 (PgR) cases of primary breast cancer. The slides were independently re-assessed by 22 pathologists from nine hospitals. A simple assessment protocol was used with a cut-off value > 10% stained nuclei, regardless of staining intensity. The concordance was high, with a kappa value of 0.78 for ER and 0.72 for PgR. Since some of the 22 pathologists considered one or several sections to be inevaluable, the evaluation comprised three groups (< or = 10% vs. > 10% vs. not evaluated). Although the percentage of positive cases varied among the 22 pathologists (ER: 57%-79%; PgR: 41%-66%), the overall concordance was good for both ER and PgR (kappa = 0.78 and 0.72, respectively). A further improvement in concordance should be possible to achieve through training.     

Interobserver variability between general and expert pathologists during the histopathological assessment of large-core needle and open biopsies of non-palpable breast lesions

The purpose of this study was to assess whether general pathologists are able to make as accurate and reproducible a diagnosis on large-core needle biopsies as on open breast biopsy specimens. A total of 688 patients underwent a stereotactic large-core (14G) needle biopsy and subsequent surgical excision of 718 non-palpable breast lesions. Forty-two pathologists from 10 departments of pathology (generalists) made a diagnosis on both the needle and open biopsy specimens. Afterwards, three pathologists and two radiologists with extensive experience in breast pathology (experts) diagnosed all of the biopsy specimens. The general pathologists made a similar histological diagnosis as the experts in 632 (88%) of the needle biopsies and 649 (90%) of the open biopsy specimens. Accordingly, the interobserver agreement for the diagnosis of large-core needle biopsies between the general and experts pathologists was excellent (kappa 0.83) and not significantly different from the interobserver agreement for the diagnosis of open breast biopsies (kappa 0.86). However, many inconsistencies were observed in the category of borderline lesions: only 24% of the large-core needle biopsies and 43% of the open biopsies with an expert diagnosis of 'borderline' were diagnosed similarly by the general pathologists. Additionally, the risk of benign/malignant inconsistencies between general pathologists and experts was approximately 1 in 55 for both needle and open biopsies.     

Mature B-cell lymphoma/leukemia in children and adolescents: intergroup pathologist consensus with the revised European-American Lymphoma Classification.

Background:The Revised European–American Lymphoma(R.E.A.L.) Classification criteria were evaluated in the internationalprotocol FAB LMB 96 Treatment of Mature B-Cell Lymphoma/Leukemia: A SFOP LMB96/CCG-5961/UKCCSG NHL 9600 Cooperative Study. This includes B-lineagelymphomas: Burkitt's lymphoma (including ALL-L3); high-grade B-cell lymphoma,Burkitt-like; diffuse large B-cell lymphoma (excluding anaplastic large cellKi-1 lymphoma). Patients and methods:Cases were independently reviewed by eighthematopathologists from the three cooperative national groups (two SFOP, twoCCG, four UKCCSG), without prior discussion of classification criteria orguidelines for case rejection. Consensus diagnosis was determined by eachnational cooperative group, and final consensus diagnosis established when atleast two national consensus diagnoses were in agreement, or following groupagreement at a multiheaded microscope. Results:Two hundred eight cases were reviewed, with finalconsensus diagnosis established in two hundred three. The percent agreementof each group's national consensus diagnosis with final consensus diagnosiswas 86%, 86% and 71%. The percent agreement of thegroup's national consensus diagnosis with final consensus diagnosis forBurkitt's and diffuse large B-cell lymphoma were 88% and 80%,respectively, but only 42% for Burkitt-like lymphoma. Conclusions:International panel review of mature B-celllymphoma/leukemia in children and adolescents highlighted difficulties insubclassification, particularly with Burkitt-like, which is a 'provisionalentity' in the R.E.A.L. Classification. The absence of previous discussion ofclassification and guidelines for case rejection may in part explain thediscrepancy between pathologists. These results underline that morphology mayneed to be complemented by other studies, such as molecular genetics andcytogenetics, to discriminate between the mature B-cell lymphomas.