Biomedical subjects

M J Brisco

Publications and source records attributed to M J Brisco.

23 records · Page 2Linked to original sources

PubMed1991

Detection and quantitation of neoplastic cells in acute lymphoblastic leukaemia, by use of the polymerase chain reaction.

We report a simple and robust method for sensitive quantitation of leukaemic cells in acute lymphocytic leukaemia. Chain determining region 3 (CDR3) of the immunoglobulin heavy chain gene is a precise genetic marker for a patient's leukaemic clone. Quantitation of the leukaemic lymphocytes was achieved by use of the polymerase chain reaction to detect CDR3 at limiting dilution of DNA samples. Five patients were studied and high levels (1 in 1 to 1 in 10) of leukaemic cells were detected at diagnosis or relapse. Leukaemic cells were detected in remission marrows from three patients, at levels of 1 in 1000 to 1 in 100,000. All five patients showed a 1000 to 100,000-fold reduction in the levels of leukaemic cells after induction therapy. This technique should prove useful for monitoring therapy and may help predict outcome.

Adult↗

PubMed1990

Monoclonality in B-lymphoproliferative disorders detected at the DNA level.

A new method was developed for detection of monoclonality at the DNA level in B-lymphoproliferative disease using the polymerase chain reaction and consensus primers for the V and J regions of the immunoglobulin gene. Monoclonality was detected in DNA from 15 of 15 clonal B-lymphoblastoid cell lines and from 19 of 23 cases of B-lymphocyte neoplasia, but not from any of 16 normal T-lymphocyte clones, 9 cases of T-lymphocyte neoplasia, 20 samples of polyclonal peripheral blood lymphocytes, and 8 cases of reactive lymphadenopathy. This method for detection of monoclonality is likely to be of routine value in diagnosis owing to its simplicity, speed, and versatility.

B-Lymphocytes↗

PubMed1990

Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B-lymphocyte clones in a polyclonal population.

A method has been developed to use the polymerase chain reaction to amplify and sequence the chain determining region 3 (CDR 3) of the human immunoglobulin heavy-chain gene, and to use the sequence as a marker for rare neoplastic B lymphocytes. Consensus primers for the Variable and Joining regions of the gene were constructed and shown to enable efficient amplification, directed cloning, and sequencing of CDR 3. Using leukaemic cell line PFMC as a test system, CDR 3 was sequenced, specific primers synthesized, and PFMC DNA was detected down to a dilution of 1:1300 in DNA from normal lymphocytes. This strategy should be useful for monitoring therapy and detecting early disease relapse in B lymphoproliferative disease.

B-Lymphocytes↗

PubMed1990

Monoclonality in B cell lymphoma detected in paraffin wax embedded sections using the polymerase chain reaction.

The polymerase chain reaction (PCR) was used to develop a simple technique for detecting monoclonality at the DNA level in B lymphocyte populations in formalin fixed, paraffin wax embedded material. Sections were dewaxed and dehydrated, and the DNA was extracted by boiling in water for 45 minutes. A semi-nested PCR was performed to amplify the V-D-J region of the immunoglobulin heavy chain gene. The product was electrophoresed and viewed under ultraviolet light after ethidium bromide staining. Specimens from 26 B cell lymphomas produced a monoclonal band in 24 cases and no amplification in two cases; monoclonality was specific for this disorder. Specimens from seven T cell lymphomas produced no amplification; specimens from nine reactive nodes produced a broad smear of polyclonal material; and specimens from 12 cases of carcinoma produced either no amplification or polyclonal material. As detection of monoclonality is strongly suggestive of neoplastic disease, this technique is likely to be of value in routine diagnosis, because of its speed, simplicity, and applicability to fixed, embedded material.

DNA, Neoplasm↗

PubMed1989

Southern bean mosaic virus RNA remains associated with swollen virions during translation in wheat germ cell-free extracts.

L-[35S]Methionine-labeled translation complexes were prepared by incubating either swollen intact southern bean mosaic virus (SBMV) particles or unencapsidated SBMV RNA in a wheat germ extract. The complexes were analysed by sucrose gradient centrifugation and by electron microscopy and dot blot hybridization of fractions from these gradients. In these complexes, 80 S ribosomes appeared to be associated with intact or near intact particles, suggesting that SBMV particles disassemble only after their RNA has initiated translation. This is in contrast to some other isometric viruses, such as turnip yellow mosaic virus, which appear to release their RNA rapidly prior to translation.

Cell-Free System↗