ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

The utility of polymerase-mediated assays in the detection of thet(11;14) involving thebcl-1 major translocation cluster (bcl-1 MTC) was evaluated by analyzing DNA from 33 patients with mantle cell lymphoma, 14 patients with other non-Hodgkin s lymphomas, and five patients with reactive lymphoid hyperplasia. The polymerase chain reaction (PCR) assay was performed using a consensus immunoglobin heavy-chain joining region primer in conjunction with a chromosome 11 specific oligonucleotide primer flanking the translocation site. The sensitivity and specificity of the assay were confirmed by correlation of the (PCR) assay data with restriction analysis. Rearrangements at thebcl-1 MTC were detected in 13 (39%) of 33 cases of mantle cell lymphoma by PCR and in 13 (48%) of 27 cases by restriction analysis. Amplicons were detectable by PCR in 85% (11 of 13) of the cases shown to bebcl-1 rearranged by restriction analysis. Failure to detect amplification products in DNA samples from non-mantle cell lymphomas and reactive follicular hyperplasia further confirmed the specificity of the assay. Sequential hybridization of the PCR products with oligonucleotide probes 3′ to thebcl-1 MTC primer revealed that the breakpoints in thebcl-1 MTC were clustered around anSstI restriction site over a range of 170 base pairs. The study demonstrates that PCR-mediated assay for the detection of thet(l1;14) at thebcl-1 MTC is specific and sensitive and can be used as an adjunct to restriction analysis in routine diagnostics.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

To determine efficiently the clonality of B-cell lymphoproliferative disorders, we modified an immunoglobulin heavy-chain (IGH) gene rearrangement polymerase chain reaction (PCR) assay that requires only a single primer germline variable (VH) and joining (JH) pair and does not involve nested priming, blot hybridization, radioactivity, or sequencing of the amplified PCR product. This simple PCR technique enabled detection of IGH gene rearrangements in as little as 10 pg (one cell equivalent) of DNA or when the clonal-to-polyclonal B-cell ratio was experimentally set at 1:1000. We detected IGH gene rearrangements in 83.5% (71 of 85) of clonal B-cell processes, a sensitivity approaching that of more cumbersome multiple primer and nested primer assays. Moreover, this technique is equally effective with fixed tissues, either B5 or formalin, and can be performed on minute samples, histologic sections, fine-needle aspirates, or cerebrospinal fluids. When compared with conventional Southern blot analysis using a genomic JHprobe, the PCR assay demonstrated IGH gene rearrangements in 82% (37 of 45) of B-cell processes positive by Southern blot. No false-positive results were observed in 29 negative control tissues. We now use IGH gene PCR routinely in our laboratory for the detection of clonal B-cells in virtually any tissue sample as an aid in early diagnosis, staging, and monitoring, and the Southern blot procedure is reserved for only a minority of diagnostic cases.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

We evaluated four polymerase chain reaction (PCR) methods for their efficiency in detecting monoclonality in a well-characterized panel of frozen and paraffin-embedded B-cell lymphoid proliferations. These approaches (referred to as FR3, FR3A, FR2, and FRI) are based on amplification of rearranged immunoglobulin heavy chain genes, using primers recognizing framework regions I, II, or III. FR3, FR3A and FR2 approaches reproducibly detected monoclonality in 51%, 72%, and 67% of DNAs from frozen lymphomas, respectively. No false-positives were observed. The combination of FR2 and FR3A methods raised the figure to 85%. Comparable results were obtained using paraffin-embedded lymphomas. Reproducibility of FRI approach was unsatisfactory. The efficiency of all PCR approaches varied depending on lymphoma type. The highest detection rate was in small/intermediate cell and the lowest in centro-follicular lymphomas. Limiting dilution assays showed that PCR methods were able to detect monoclonal B-cell DNA representing 5% of nonlymphoid and 20% of polyclonal B-cell DNA. A diagnostic protocol may include quick and cost-effective PCR screening, particularly in cases of undetermined small cell lymphoid proliferations observed in fine needle aspirates or endoscopic biopsies. This would also reduce call-up of patients to obtain unfixed biopsies.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

Cytologic evaluation of lymph node fine-needle aspirates and serous effusions is a rapid and useful means for establishing the diagnosis of a variety of lymphoproliferative disorders. However, in some instances, cytologic findings are not sufficient to establish a diagnosis of lymphoma, thus necessitating the use of ancillary procedures, the most frequent of which is immunophenotyping. In this respect, the usefulness of molecular markers, such as clonal immunoglobulin gene rearrangements or chromosomal translocations, have been less well evaluated. Follicular lymphoma constitutes an interesting disease for such a study because these tumors possess characteristic histopathologic features and contain two potential molecular markers, that is, a clonal immunoglobulin gene rearrangement and abcl-2 gene translocation [t (14; 18)]. In the present study, we evaluated, retrospectively, the cytologic material from four lymph node fine-needle aspirates and one pleural effusion of five patients with biopsy-proven follicular lymphoma. In four of the cases, definitive diagnosis of lymphoma had not been possible solely from cytologic evaluation. DNA was isolated from archival air-dried samples present on glass slides and amplified by the polymerase chain reaction (PCR) for detection of either a clonal immunoglobulin heavy chain gene rearrangement orbcl-2 translocation (major breakpoint region). An immunoglobulin heavy-chain gene rearrangement was detected in four of five patients, and two patients had thebcl-2 translocation by PCR. The effusion case was identical by gel electrophoresis with product amplified from a lymph node biopsy of the same patient and DNA extracted directly from fresh pleural effusion cells. The results indicate that PCR may be a useful ancillary procedure in evaluation of patients with follicular lymphoma when the diagnosis cannot be established by morphology alone. This technique does not require any additional cytologic material because the sample present on the original air-dried glass slide can be used as the source of the required DNA.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

Diagnosis of gastric malignant lymphoma remains a challenge, especially when the tissue source is endoscopic biopsy specimens. Once an atypical lymphoid infiltrate is found, demonstration of clonality is the key to establishing a diagnosis of the disease. For this purpose, we evaluated the usefulness of immunohistochemistry, in situ hybridization, and polymerase chain reaction (PCR) using formalin-fixed, paraffin-embedded endoscopic materials from 20 patients with B-cell malignant lymphomas. Template DNA for PCR was obtained by microdissecting Giemsa-stained sections using a serial hematoxylin and eosin section as a guide. Clonal rearrangement bands were demonstrated in 15 of 20 cases (75%) by PCR, whereas expression of monotypic light-chain mRNA was detected in seven of 20 (35%) by in situ hybridization and monotypic light-chain restriction in four of 20 (20%) by conventional immunohistochemistry. Although less sensitive than PCR, in situ hybridization was useful for localizing the expression of target mRN As with cellular accuracy and with low background staining. In addition, two cases were found to be monoclonal only by in situ hybridization, and not by PCR. The results showed that clonal proliferation is detected with the greatest sensitivity with PCR using small routinely processed biopsy specimens and that a difficulty with the PCR method in terms of cellular localization was partially overcome using a microdissection procedure that provided at least tissue-level accuracy.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

We report a case of a 21-year-old woman with hematopoietic. immunological, and congenital dysmorphic abnormalities, who died following rapidly progressive, disseminated Epstein-Barr virus (EBV)-associated lymphoproliferative disease (LPD). Polymerase chain reaction (PCR) amplification of formalin-fixed paraffin-embedded tissue showed differences in the clonality of each separate lymphoproliferative lesion examined, as determined by immunoglobulin heavy chain (IgH) gene rearrangement. PCR analysis also demonstrated that all lesions contained EBV genome. Since DNA had been extracted from paraffin blocks, a direct comparison of morphology and clonality could be made in each individual lesion. The evidence from this study indicates that the monoclonal tumors arose de novo in multiple sites and that the polyclonal background observed in some lesions reflected a substantial concomitant inflammatory response.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

Carcinoid tumor of the kidney is a rare neoplasm of uncertain histogenesis. Attempts to elucidate its cell of origin have been made, but there is a lack of experimental proof. We present a case of primary renal carcinoid tumor with a characteristic molecular abnormality and discuss its histogenetic implications. Histologic, immunohisto-chemical, and electron microscopic analyses revealed features typical of carcinoid tumor, and DNA flow cytometric analysis showed diploid pattern. Molecular genetic studies of informative WT1, p53, and 3p21 loci revealed loss of heterozygosity only at the D3F15S2 locus (3p21 telomeric). The similarity between the molecular abnormality in the present case and that in most renal cell carcinomas suggests a possible common genetic event in the genesis of these neoplasms.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID

摘要:

Many human cancers present deletions of the short arm of chromosome 17, which includes the TP53 locus. We detected a new polymorphism in intron 2 of the TP53 gene using PCR-SSCP and used this polymorphic site as a marker to detect loss of heterozygosity in 135 human tumors (73 soft tissue sarcomas, and 48 colorectal and 14 bladder carcinomas). Heterozygosity for this site was 41.5% in this study group and tumor-specific loss of alleles occurred in 43% of informative cases. Allelic losses were more frequently detected at this site than at that in which restriction fragment length polymorphism (RFLP) is located, as detected by the pHp53B probe. It is concluded that this novel approach has several advantages, including detection of a high incidence of informative cases and minimal tissue requirements.

ISSN:1052-9551    

出版商:OVID    

年代:1995

数据来源: OVID