Polymerase chain reaction-based monitoring techniques for the detection of insecticide resistance-associated point mutations and their potential applications

1995 ◽  
Vol 43 (3) ◽  
pp. 195-200 ◽  
Author(s):  
Richard Ffrench-Constant ◽  
Kate Aronstein ◽  
Nicola Anthony ◽  
Christine Coustau
Keyword(s):  
Polymerase Chain Reaction ◽  
Insecticide Resistance ◽  
Point Mutations ◽  
Chain Reaction ◽  
Monitoring Techniques ◽  
Potential Applications ◽  
Polymerase Chain

Norwalk-like Virus Sequences Detected by Reverse Transcription-Polymerase Chain Reaction in Mineral Waters Imported into or Bottled in Switzerland

2000 ◽  
Vol 63 (11) ◽  
pp. 1576-1582 ◽  
Author(s):  
CHRISTIAN BEURET ◽  
DOROTHE KOHLER ◽  
THOMAS LÜTHI
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Carbonic Acid ◽  
Point Mutations ◽  
Chemical Properties ◽  
The United States ◽  
Mineral Waters ◽  
Nucleotide Sequence Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

Norwalk-like viruses (NLVs) is a genus belonging to the Caliciviridae. NLVs are transmitted by the fecal-oral and the aerosol route and are the most common cause of outbreaks of nonbacterial gastroenteritis. NLVs are responsible for an estimated 67% of all illnesses caused by known foodborne pathogens and for 96% of nonbacterial gastroenteritis in the United States. Many outbreaks could be associated with the consumption of primarily or secondarily contaminated foods. To our knowledge, no epidemic arising from contaminated mineral water has been reported. We investigated the presence of NLV sequences in 63 mineral waters of 29 different brands that were imported into or bottled in Switzerland. NLV sequences were detected in 21 mineral waters by reverse transcription-seminested polymerase chain reaction. Specimens of two NLV genogroups (gg), gg I and gg II, were randomly present in the contaminated samples. The presence of NLV sequences could not be correlated either with bottle characteristics or with chemical properties like mineralization, pH, or the presence of carbonic acid. Nucleotide sequence analysis of 12 NLV-positive samples revealed several point mutations. All isolated NLV gg I strains have a similarity of 70 to 87% with the common Desert Shield virus (UO4469), and all isolated NLV gg II strains have a similarity of 89 to 93% with the Camberwell virus (U46500). Possible reasons for the presence of NLV sequences in mineral waters are discussed.

scholarly journals Precision of genotyping of Epstein-Barr virus by polymerase chain reaction using three gene loci (EBNA-2, EBNA-3C, and EBER): predominance of type A virus associated with Hodgkin's disease [published erratum appears in Blood 1993 Oct 1;82(7):2268]

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3372-3381 ◽  
Author(s):  
JC Lin ◽  
SC Lin ◽  
BK De ◽  
WC Chan ◽  
BL Evatt ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cell Lines ◽  
Epstein Barr Virus ◽  
Point Mutations ◽  
Type A ◽  
Type B ◽  
Chain Reaction ◽  
Barr Virus ◽  
Polymerase Chain ◽  
Epstein Barr

Abstract To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin's disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31% with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.

scholarly journals Precision of genotyping of Epstein-Barr virus by polymerase chain reaction using three gene loci (EBNA-2, EBNA-3C, and EBER): predominance of type A virus associated with Hodgkin's disease [published erratum appears in Blood 1993 Oct 1;82(7):2268]

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3372-3381 ◽  
Author(s):  
JC Lin ◽  
SC Lin ◽  
BK De ◽  
WC Chan ◽  
BL Evatt ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cell Lines ◽  
Epstein Barr Virus ◽  
Point Mutations ◽  
Type A ◽  
Type B ◽  
Chain Reaction ◽  
Barr Virus ◽  
Polymerase Chain ◽  
Epstein Barr

To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin's disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31% with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.

scholarly journals Inactivation of the p15 gene in children with acute lymphoblastic leukemia

2003 ◽  
Vol 121 (5) ◽  
pp. 203-206 ◽  
Author(s):  
Rosana Cipolotti ◽  
José Alexandre Rodrigues Lemos ◽  
Ricardo Defavery ◽  
Carlos Alberto Scrideli ◽  
Amaury Lellis Dal Fabbro ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Point Mutations ◽  
Prognostic Indicators ◽  
Chain Reaction ◽  
Exon 2 ◽  
Polymerase Chain ◽  
P15 Gene

CONTEXT: Tumor suppressor genes act on the control of cell cycle progression. In pediatric neoplasias, some of these genes may be considered to be markers for diagnosis or relapse, thus probably representing prognostic indicators. OBJECTIVE: To study the inactivation of the p15 gene in children with acute lymphoblastic leukemia. TYPE OF STUDY: Retrospective study. SETTING: Laboratory of Molecular Biology, Department of Pediatrics, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo. PARTICIPANTS: Eighty-three children and adolescents with acute lymphoblastic leukemia were studied, with the examination of 83 bone marrow samples obtained at diagnosis, four obtained also during relapse, and two cerebrospinal fluid samples obtained from two cases of isolated relapse in the central nervous system. MAIN MEASUREMENTS: Homologous deletion of the p15 gene by multiplex polymerase chain reaction, and screening for point mutations by polymerase chain reaction/single-strand conformational polymorphism. RESULTS: Deletion of exon 2 of the p15 gene was observed in 15 children, including one case in which deletion was only verified during isolated central nervous system relapse. No case of exon 1 deletion, or that was suggestive of point mutations, was observed and no association between p15 gene inactivation and classic risk factors was established. CONCLUSION: According to the literature, inactivation of the p15 gene by deletion of exon 2 in acute lymphoblastic leukemia found in the population studied would be considered to be a molecular marker for diagnosis or relapse. However, no correlation between p15 gene deletion and clinical prognostic indicators was observed.

scholarly journals Evidence of Genetic Diversity Underlying Rh D−, Weak D (Du), and Partial D Phenotypes as Determined by Multiplex Polymerase Chain Reaction Analysis of the RHD Gene

Blood ◽  
1997 ◽  
Vol 89 (7) ◽  
pp. 2568-2577 ◽  
Author(s):  
Neil D. Avent ◽  
Peter G. Martin ◽  
Sylvia S. Armstrong-Fisher ◽  
Wendy Liu ◽  
Kirstin M. Finning ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Basis ◽  
Gene Deletion ◽  
Stop Codon ◽  
Point Mutations ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Multiplex Pcr Assay ◽  
Polymerase Chain

Abstract The human blood group Rh antigens are expressed by proteins encoded by a pair of highly homologous genes located at chromosome 1p34-36. One of the genes (RHCE ) encodes Rh CcEe antigens, while the other (RHD) the D antigen. Point mutations in the RHCE gene generate the C/c and E/e polymorphisms, while it has been shown that an RHD gene deletion can generate the D-negative phenotype. We have analyzed intron 4 of the RHCE and RHD genes and have defined the site of an RHD-specific deletion located in this intron. Using a multiplex RHD typing assay, which combines a reverse polymerase chain reaction (PCR) primer, which straddles this RHD-specific sequence, and a pair of primers located in exon 10 of the RHD gene, we have analyzed 357 different genomic DNA samples derived from individuals expressing D+, D−, weak D, and partial D phenotypes. Of these, we have noted a significant discordance with our multiplex PCR assay in the D− phenotypes dCcee and dccEe (which have been previously described) and weak D phenotypes. Our results suggest that in five serologically D− individuals we have identified an apparently intact RHD gene. Sequence analysis of transcripts obtained from one of these individuals (of phenotype dCCee) illustrates the presence of full-length RHD transcripts, which have a point mutation at nucleotide 121 (C → T), which generates an in-frame stop codon (Gln41Stop). Thus, we describe a different molecular basis for generating the D− phenotype to the complete RHD gene deletion described previously. We also show that there are discordances with serotype and the multiplex assay in weak D and partial D phenotypes, indicating that the underlying molecular basis can be heterogeneous. Existing Rh D PCR assays assume the complete absence of the RHD gene in D− phenotypes. We describe a different molecular basis for generating the D− phenotype to the complete RHD gene deletion described previously.

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