Detection of chromosomal translocation t(14;18) within the minor cluster region of bcl-2 by polymerase chain reaction and direct genomic sequencing of the enzymatically amplified DNA in follicular lymphomas

A majority of t(14;18) translocations have been shown to cluster at one of two sites on chromosome 18, called the major breakpoint region (mbr) or the minor cluster region, (mcr), which map within or flanking the bcl-2 proto-oncogene, respectively. We have determined the nucleotide sequence for a portion of the mcr, and constructed oligonucleotides that were used to perform the polymerase chain reaction (PCR) in conjunction with universal immunoglobulin primers to specifically amplify t(14;18) breakpoints in DNA obtained from follicular lymphomas. Eight of ten breakpoints that were detectable on Southern blots using DNA probes for the mcr could be detected due to specific amplification by the PCR technique using an mcr-specific primer. Direct nucleotide sequencing of the enzymatically amplified DNAs showed that the breakpoints clustered within a 500 nucleotide region, and five occurred within three nucleotides of each other. These data show a remarkable clustering of some t(14;18) breakpoints at a site on chromosome 18, at least a 30-kb distance from the bcl-2 gene. Our findings also indicate that mcr-specific primers may be used in conjunction with previously described mbr-specific primers in a highly sensitive DNA amplification technique to detect a large fraction of t(14;18) breakpoints.

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Detection of chromosomal translocation t(14;18) within the minor cluster region of bcl-2 by polymerase chain reaction and direct genomic sequencing of the enzymatically amplified DNA in follicular lymphomas

Blood ◽

10.1182/blood.v73.7.1759.1759 ◽

1989 ◽

Vol 73 (7) ◽

pp. 1759-1762 ◽

Cited By ~ 154

Author(s):

BY Ngan ◽

J Nourse ◽

ML Cleary

Keyword(s):

Polymerase Chain Reaction ◽

Large Fraction ◽

Dna Amplification ◽

Nucleotide Sequencing ◽

Chromosome 18 ◽

Chain Reaction ◽

Specific Primers ◽

Cluster Region ◽

Polymerase Chain ◽

Follicular Lymphomas

Abstract A majority of t(14;18) translocations have been shown to cluster at one of two sites on chromosome 18, called the major breakpoint region (mbr) or the minor cluster region, (mcr), which map within or flanking the bcl-2 proto-oncogene, respectively. We have determined the nucleotide sequence for a portion of the mcr, and constructed oligonucleotides that were used to perform the polymerase chain reaction (PCR) in conjunction with universal immunoglobulin primers to specifically amplify t(14;18) breakpoints in DNA obtained from follicular lymphomas. Eight of ten breakpoints that were detectable on Southern blots using DNA probes for the mcr could be detected due to specific amplification by the PCR technique using an mcr-specific primer. Direct nucleotide sequencing of the enzymatically amplified DNAs showed that the breakpoints clustered within a 500 nucleotide region, and five occurred within three nucleotides of each other. These data show a remarkable clustering of some t(14;18) breakpoints at a site on chromosome 18, at least a 30-kb distance from the bcl-2 gene. Our findings also indicate that mcr-specific primers may be used in conjunction with previously described mbr-specific primers in a highly sensitive DNA amplification technique to detect a large fraction of t(14;18) breakpoints.

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High-resolution HLA-DQB1 typing using the polymerase chain reaction and sequence-specific primers

Tissue Antigens ◽

10.1111/j.1399-0039.1997.tb02936.x ◽

1997 ◽

Vol 50 (6) ◽

pp. 688-692 ◽

Cited By ~ 14

Author(s):

C.G. Mullighan ◽

M. Bunce ◽

K.I. Welsh

Keyword(s):

Polymerase Chain Reaction ◽

High Resolution ◽

Chain Reaction ◽

Specific Primers ◽

Polymerase Chain

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PCR BY THERMAL CONVECTION

Modern Physics Letters B ◽

10.1142/s0217984904007049 ◽

2004 ◽

Vol 18 (16) ◽

pp. 775-784 ◽

Cited By ~ 37

Author(s):

DIETER BRAUN

Keyword(s):

Polymerase Chain Reaction ◽

Thermal Convection ◽

Dna Amplification ◽

Reaction Vessel ◽

Cold Region ◽

Chain Reaction ◽

Heating And Cooling ◽

Highly Sensitive ◽

Specific Amplification ◽

Polymerase Chain

The Polymerase Chain Reaction (PCR) allows for highly sensitive and specific amplification of DNA. It is the backbone of many genetic experiments and tests. Recently, three labs independently uncovered a novel and simple way to perform a PCR reaction. Instead of repetitive heating and cooling, a temperature gradient across the reaction vessel drives thermal convection. By convection, the reaction liquid circulates between hot and cold regions of the chamber. The convection triggers DNA amplification as the DNA melts into two single strands in the hot region and replicates into twice the amount in the cold region. The amplification progresses exponentially as the convection moves on. We review the characteristics of the different approaches and show the benefits and prospects of the method.

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A rapid molecular method (polymerase chain reaction with sequence-specific primers) to genotype for ABO blood group and secretor status and its potential for organ transplants

Tissue Antigens ◽

10.1111/j.1399-0039.1997.tb02902.x ◽

1997 ◽

Vol 50 (5) ◽

pp. 475-483 ◽

Cited By ~ 27

Author(s):

J. Procter ◽

J. Crawford ◽

M. Bunce ◽

K.I. Welsh

Keyword(s):

Polymerase Chain Reaction ◽

Blood Group ◽

Abo Blood Group ◽

Organ Transplants ◽

Molecular Method ◽

Chain Reaction ◽

Specific Primers ◽

Secretor Status ◽

Polymerase Chain

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Genetic diversity and phylogeny analysis of Azolla based on DNA amplification by arbitrary primers

Genome ◽

10.1139/g93-092 ◽

1993 ◽

Vol 36 (4) ◽

pp. 686-693 ◽

Cited By ~ 23

Author(s):

Benoit Van Coppenolle ◽

Iwao Watanabe ◽

Charles Van Hove ◽

Gerard Second ◽

Ning Huang ◽

...

Keyword(s):

Principal Component Analysis ◽

Polymerase Chain Reaction ◽

Principal Component ◽

Dna Amplification ◽

Component Analysis ◽

Genetic Distances ◽

Group Method ◽

Chain Reaction ◽

Arbitrary Primers ◽

Polymerase Chain

The polymerase chain reaction was used to amplify random sequences of DNA from 25 accessions of Azolla to evaluate the usefulness of this technique for identification and phylogenetic analysis of this aquatic fern. Accessions were selected to represent all known species within the genus Azolla and to encompass the worldwide distribution of the fern. Primers of 10 nucleotides with 70% G + C content were used to generate randomly amplified polymorphic DNA from the symbiotic Azolla–Anabaena complex. Twenty-two primers were used and each primer gave 4–10 bands of different molecular weights for each accession. Bands were scored as present or absent for each accession and variation among accessions was quantified using Nei's genetic distances. A dendrogram summarizing phenetic relationships among the 25 accessions was generated using the unweighted pair-group method with arithmetic mean. Principal component analysis was also used to evaluate genetic similarities. Three distinct groups were identified: group 1 contains five species, group 2 contains the pinnata species, and group 3 contains the nilotica species. The analysis demonstrates that the major groups of Azolla species can be easily distinguished from one an other and, in addition, that closely related accessions within species can be identified. We further found that using 10 primers, a phylogeny that is essentially the same as that derived from 22 primers can be constructed. Our results suggest that total DNA extracted from the Azolla–Anabaena symbionts is useful for classification and phylogenetic studies of Azolla.Key words: Azolla–Anabaena symbiosis, genetic distances, polymerase chain reaction, principal component analysis.

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Detection of microchimerism after allogeneic blood transfusion using nested polymerase chain reaction amplification with sequence-specific primers (PCR-SSP): A cautionary tale

Transfusion Medicine Reviews ◽

10.1016/s0887-7963(99)80094-7 ◽

1999 ◽

Vol 13 (1) ◽

pp. 69

Author(s):

Sunny Dzik

Keyword(s):

Polymerase Chain Reaction ◽

Blood Transfusion ◽

Polymerase Chain Reaction Amplification ◽

Allogeneic Blood ◽

Cautionary Tale ◽

Chain Reaction ◽

Specific Primers ◽

Nested Polymerase Chain Reaction ◽

Reaction Amplification ◽

Polymerase Chain

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HTLV-I-induced lymphoma mimicking Hodgkin's disease. Diagnosis by polymerase chain reaction amplification of specific HTLV-I sequences in tumor DNA

Blood ◽

10.1182/blood.v71.4.1027.1027 ◽

1988 ◽

Vol 71 (4) ◽

pp. 1027-1032 ◽

Cited By ~ 87

Author(s):

DB Duggan ◽

GD Ehrlich ◽

FP Davey ◽

S Kwok ◽

J Sninsky ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Hodgkin's Disease ◽

Polymerase Chain Reaction Amplification ◽

Hodgkin’S Disease ◽

Dna Amplification ◽

Disease Diagnosis ◽

Lymphoproliferative Disease ◽

Enzyme Linked Immunosorbent Assay ◽

Chain Reaction ◽

Polymerase Chain

Abstract A patient with a localized HTLV-I-associated lymphoproliferative disease that was misdiagnosed as Hodgkin's disease is presented. The patient's serum was negative for HTLV-I antibodies by enzyme-linked immunosorbent assay (ELISA), Western blot, and radioimmunoprecipitation. Tumor tissue DNA was negative for HTLV-I by Southern blotting but was positive for distinct HTLV-I sequences when subjected to DNA amplification using the polymerase chain reaction. We conclude that the clinical and pathologic diagnosis of HTLV-I-related lymphoma can be difficult and can be confused with Hodgkin's disease. Extremely sensitive molecular biological techniques may be required to establish a diagnosis of HTLV-I-induced lymphoma.

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