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Multiplex polymerase chain reaction

About: Multiplex polymerase chain reaction is a research topic. Over the lifetime, 6409 publications have been published within this topic receiving 221244 citations.


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Journal Article
01 Jan 1994-Leukemia
TL;DR: A rapid and simple polymerase chain reaction (PCR) method is described that is capable of identifying any of the BCR-ABL transcripts that have yet been described in chronic myeloid or acute lymphoblastic leukaemia.
Abstract: A rapid and simple polymerase chain reaction (PCR) method is described that is capable of identifying any of the BCR-ABL transcripts that have yet been described in chronic myeloid or acute lymphoblastic leukaemia. Randomly primed cDNA is synthesized from leucocyte RNA and amplified in a single reaction containing four oligonucleotide primers (multiplex PCR). Different size products are generated from ela2 (p190) and b3a2 or b2a2 (p210) BCR-ABL transcripts which are readily and unambiguously distinguishable after agarose gel electrophoresis without the need for either nested PCR or hybridization. Chronic myeloid leukaemia cells are readily detectable even when diluted 1 in 1000 with normal blood. Samples which do not have BCR-ABL rearrangements produce a single band derived from the normal BCR gene, and the presence of this band controls for adequate RNA and cDNA preparation. Using this assay we have detected BCR-ABL transcripts in a variety of haematological disorders.

284 citations

Journal ArticleDOI
01 Feb 2007-Leukemia
TL;DR: B-cell clonality was detected by combined use of the IGH and IGK multiplex PCR assays in all 260 definitive cases of B-cell chronic lymphocytic leukemia, mantle cell lymphoma, marginal zone lymphoma and follicular lymphoma.
Abstract: Polymerase chain reaction (PCR) assessment of clonal immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements is an important diagnostic tool in mature B-cell neoplasms. However, lack of standardized PCR protocols resulting in a high level of false negativity has hampered comparability of data in previous clonality studies. In order to address these problems, 22 European laboratories investigated the Ig/TCR rearrangement patterns as well as t(14;18) and t(11;14) translocations of 369 B-cell malignancies belonging to five WHO-defined entities using the standardized BIOMED-2 multiplex PCR tubes accompanied by international pathology panel review. B-cell clonality was detected by combined use of the IGH and IGK multiplex PCR assays in all 260 definitive cases of B-cell chronic lymphocytic leukemia (n=56), mantle cell lymphoma (n=54), marginal zone lymphoma (n=41) and follicular lymphoma (n=109). Two of 109 cases of diffuse large B-cell lymphoma showed no detectable clonal marker. The use of these techniques to assign cell lineage should be treated with caution as additional clonal TCR gene rearrangements were frequently detected in all disease categories. Our study indicates that the BIOMED-2 multiplex PCR assays provide a powerful strategy for clonality assessment in B-cell malignancies resulting in high Ig clonality detection rates particularly when IGH and IGK strategies are combined.

282 citations

Journal ArticleDOI
TL;DR: Analysis of the sometimes complex characteristics of mul?
Abstract: with the potential to be phylogenetically informative (e.g., sequence variation, vari? ation within gene clusters, and number of gene clusters [Koop et al., 1989; Bartels et al., 1993; Cartwright et al., 1993; Pendleton et al., 1993]). Nevertheless, this complexity is sometimes difficult to interpret in a phy? logenetic context (e.g., orthologous and paralogous variation and gene conversion [Fitch, 1970; Hillis and Dixon, 1991; San? derson and Doyle, 1992]). Analysis of the sometimes complex characteristics of mul? tigene families has recently been greatly facilitated by use of the polymerase chain reaction (PCR). Using degenerate primers corresponding to highly conserved regions of homologous genes, PCR can be used to detect and identify members of these gene families in samples of genomic DNA or cDNA. The utility of such an approach has recently been demonstrated using degen? erate primers to amplify a region of Antennapedia-class homeoboxes (Murtha et al., 1991; Pendleton et al., 1993). This tech? nique, however, cannot be fully exploited until certain methodological issues have

280 citations

Journal ArticleDOI
TL;DR: A vertebrate-specific multiplexed primer set based on mitochondrial cytochrome b will be a valuable tool for identifying the blood meals of field-collected mosquitoes where people and alternative mammal hosts are present.
Abstract: To date, no polymerase chain reaction diagnostic technique exists to directly identify mammalian blood meals from mosquitoes by sized DNA fragments following agarose gel electrophoresis. We have developed a vertebrate-specific multiplexed primer set based on mitochondrial cytochrome b to identify the mammalian blood hosts of field-collected mosquitoes. Although designed for the study of African malaria vectors, the application of this tool is not restricted to this disease system. Validation of this diagnostic technique on dried anopheline and culicine field specimens collected in Zambia and Mali demonstrated that blood meals could be identified 2-7 months after collection. Time course experiments showed that host DNA was detectable in frozen mosquito abdomens 24-30 hours post-feeding. Additionally, multiple blood meals from different mammals could be detected in a single mosquito. This diagnostic assay will be a valuable tool for identifying the blood meals of field-collected mosquitoes where people and alternative mammal hosts are present.

279 citations

Journal Article
TL;DR: A two-step, multiplex polymerase chain reaction (PCR) method based on genomic regions of difference allowed for rapid differentiation of members of the Mycobacterium tuberculosis complex, making it suitable for routine laboratories and surveillance purposes.
Abstract: Differentiation of members of the Mycobacterium tuberculosis complex by conventional mycobacteriological methods is time consuming, making surveillance of species-specific disease difficult A two-step, multiplex polymerase chain reaction (PCR) method based on genomic regions of difference (RD1, RD1(mic), RD2(seal), RD4, RD9 and RD12) was developed for the differentiation of M canettii, M tuberculosis, M africanum, M microti, M pinnipedii, M caprae, M bovis and M bovis BCG The size of the respective multiplex PCR amplification products corresponded to the presence of the different M tuberculosis complex members This method allows for rapid differentiation, making it suitable for routine laboratories and surveillance purposes

279 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
2023275
2022448
2021172
2020176
2019221
2018220