scholarly journals Analysis of methicillin-resistant and methicillin-susceptibleStaphylococcus aureusby a molecular typing method based on coagulase gene polymorphisms

1995 ◽  
Vol 115 (3) ◽  
pp. 419-426 ◽  
Author(s):  
N. Kobayashi ◽  
K. Taniguchi ◽  
K. Kojima ◽  
S. Urasawa ◽  
N. Uehara ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Gene Polymorphisms ◽  
Fragment Length Polymorphism ◽  
Restriction Enzyme Digestion ◽  
Rflp Pattern ◽  
Typing Method ◽  
Methicillin Resistant ◽  
Serological Typing ◽  
Molecular Typing Method ◽  
Rflp Typing

SummaryA molecular typing method forStaphylococcus aureusbased on coagulase gene polymorphisms (coagulase gene typing) was evaluated by examining a total of 240 isolates which comprised 210 methicillin-resistantS. aureus(MRSA) and 30 methicillin-susceptibleS. aureus(MSSA) collected from a single hospital. ByAlulrestriction enzyme digestion of the PCR-amplified 3′-end region of the coagulase gene including 81-bp repeated units, the MRSA and MSSA isolates examined were divided into 6 and 12 restriction fragment length polymorphism (RFLP) patterns, respectively, whereas five patterns were commonly detected in MRSA and MSSA. MRSA isolates that showed a particular RFLP pattern were considered to be predominant in the hospital. Coagulase typing with type-specific antisera was also performed for allS. aureusisolates for comparison. Coagulase types II and VII were most frequently detected and included isolates with four and five differentAluIRFLP patterns, respectively, whereas each of the other coagulase types corresponded to a single RFLP pattern. These results indicated that RFLP typing was more discriminatory than serological typing, for typingS. aureusand demonstrated its utility in epidemiologic investigation ofS. aureusinfection in hospitals.

Localization of the Br Polymorphism on a 144 bp Exon of the GPIa Gene and Its Application in Platelet DNA Typing

1994 ◽  
Vol 71 (05) ◽  
pp. 651-654 ◽  
Author(s):  
Rainer Kalb ◽  
Sentot Santoso ◽  
Katja Unkelbach ◽  
Volker Kiefel ◽  
Christian Mueller-Eckhardt
Keyword(s):  
Genomic Organization ◽  
Fragment Length Polymorphism ◽  
Human Platelet ◽  
Dna Typing ◽  
Rflp Analysis ◽  
Serological Typing ◽  
Allele Specific ◽  
Polymerase Chain ◽  
Alloimmune Thrombocytopenia ◽  
Rflp Typing

SummaryAlloimmunization against the human platelet alloantigen system Br (HPA-5) is the second most common cause of neonatal alloimmune thrombocytopenia (NAIT) in Caucasian populations. We have recently shown that a single base polymorphism at position 1648 on platelet mRNA coding for GPIa results in an aminoacid substitution at position 505 on the mature GPIa which is associated with the two serological defined Br phenotypes.Since DNA-typing of platelet alloantigens offers possibilities for useful clinical applications, we designed genomic DNA-based restriction fragment length polymorphism (RFLP) typing for Br alloantigens. To establish this technique we analyzed the genomic organization of GPIa adjacent to the polymorphic base. Using the polymerase chain reaction (PCR) of blood cell DNA we have identified two introns (approximately 1.7 and 1.9 kb) flanking a 144 bp coding sequence of the GPIa gene encompassing the polymorphic base 1648. Based on the in- tron sequence, a PCR primer was constructed to amplify a 274 bp fragment which was used for allele-specific RFLP to determine the Br genotypes. The results of RFLP analysis using Mnll endonuclease obtained from 15 donors (2 Br37*, 2 Br^ and 11 Brb/b) correlate perfectly with serological typing by monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay.

scholarly journals A Study of HLA-Linked Genes in a Monosymptomatic Psychotic Disorder in an Indian Bengali Population

2005 ◽  
Vol 50 (5) ◽  
pp. 269-274 ◽  
Author(s):  
Monojit Debnath ◽  
Sujit K Das ◽  
Nirmal K Bera ◽  
Chitta R Nayak ◽  
Tapas K Chaudhuri
Keyword(s):  
Human Leukocyte ◽  
Delusional Disorder ◽  
Control Group ◽  
Typing Method ◽  
Hla Genes ◽  
Healthy Donors ◽  
Serological Typing ◽  
Molecular Typing Method ◽  
Paranoid Symptoms ◽  
Nested Case Control

Objective: The etiology of delusional disorder is imperfectly understood. Involvement of biological factors has long been suspected. We examined the incidence of class I human leukocyte antigens (HLAs) in patients with delusional disorder to understand the role of HLA genes and explore a possible immunogenetic etiology for delusional disorder. Methods: We used a nested case–control study design. Psychiatric reference data were available for 27 500 patients registered between 1998 and 2003. Initially, we enrolled 150 patients with delusional disorder from the India-born Bengali population, using DSM-IV diagnostic criteria. After longitudinal follow-up, 80 patients were found to have only delusional disorder, while the remaining 70 patients represented different illnesses with paranoid symptoms and were excluded. We performed serological typing on all 150 patients and applied the polymerase chain reaction–based high-resolution molecular typing method to the 80 patients with delusional disorder. Eighty healthy donors of the same ethnic background, matched for age, sex, and other socioeconomic variables, formed the control group. Results: Some of the HLA alleles were associated with delusional disorder, and the gene HLA-A*03 was found to be significantly more frequent. This gene may influence patients' susceptibility to delusional disorder. Conclusion: The study reveals important associations between HLA genes and delusional disorder. This preliminary observation may help our understanding of this disorder's genetic basis.

scholarly journals Detection of Unsuspected Cases of Nosocomial Transmission of Tuberculosis by Use of a Molecular Typing Method

2001 ◽  
Vol 33 (4) ◽  
pp. 453-459 ◽  
Author(s):  
Griselda Tudó ◽  
Julián González ◽  
Josep M. Gatell ◽  
Joan A. Caylà ◽  
Esteban Martínez ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Nosocomial Transmission ◽  
Typing Method ◽  
Molecular Typing Method

Development of a Molecular Typing Method for Pneumocystis carinii sp.f. hominis

1996 ◽  
Vol 43 (5) ◽  
pp. 34S-34S ◽  
Author(s):  
PHILIPPE M. HAUSER ◽  
DOMINIQUE S. BLANC ◽  
JACQUES BILLE ◽  
AMALIO TELENTI ◽  
PATRICK FRANCIOLI
Keyword(s):  
Molecular Typing ◽  
Pneumocystis Carinii ◽  
Typing Method ◽  
Molecular Typing Method

A PCR-RFLP typing method for adhesion molecule gene polymorphisms and allele frequencies in a normal UK population

2002 ◽  
Vol 29 (2) ◽  
pp. 109-111 ◽  
Author(s):  
R. A. Gbadegesin ◽  
C. J. Watson ◽  
S. A. Cotton ◽  
P. E. C. Brenchley ◽  
N. J. A. Webb
Keyword(s):  
Adhesion Molecule ◽  
Gene Polymorphisms ◽  
Allele Frequencies ◽  
Typing Method ◽  
Pcr Rflp ◽  
Adhesion Molecule Gene ◽  
Rflp Typing

Rapid Molecular Typing of Methicillin-ResistantStaphylococcus aureusby PCR-RFLP

2001 ◽  
Vol 22 (5) ◽  
pp. 294-298 ◽  
Author(s):  
Thomas A. Wichelhaus ◽  
Klaus-Peter Hunfeld ◽  
Boris Böddinghaus ◽  
Peter Kraiczy ◽  
Volker Schàfer ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Restriction Analysis ◽  
Protein A ◽  
Hypervariable Region ◽  
Epidemiological Surveillance ◽  
Typing Method ◽  
Rapid Pcr ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Rflp Typing

AbstractObjective:To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistantStaphylococcus aureus(MRSA) typing in routine epidemiological surveillance.Design:The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) followingHaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent tomecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype.Results:Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates.Conclusions:This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.

scholarly journals New Short Tandem Repeat-Based Molecular Typing Method for Pneumocystis jirovecii Reveals Intrahospital Transmission between Patients from Different Wards

PLoS ONE ◽  
2015 ◽  
Vol 10 (5) ◽  
pp. e0125763 ◽  
Author(s):  
Maud Gits-Muselli ◽  
Marie-Noelle Peraldi ◽  
Nathalie de Castro ◽  
Véronique Delcey ◽  
Jean Menotti ◽  
...  
Keyword(s):  
Tandem Repeat ◽  
Short Tandem Repeat ◽  
Molecular Typing ◽  
Pneumocystis Jirovecii ◽  
Typing Method ◽  
Molecular Typing Method ◽  
Short Tandem

scholarly journals Molecular strategy for ‘serotyping’ of human enteroviruses

2001 ◽  
Vol 82 (1) ◽  
pp. 79-91 ◽  
Author(s):  
Valérie Caro ◽  
Sophie Guillot ◽  
Francis Delpeyroux ◽  
Radu Crainic
Keyword(s):  
Molecular Typing ◽  
Phylogenetic Trees ◽  
Field Isolate ◽  
Human Enterovirus ◽  
Specific Information ◽  
Genomic Fragment ◽  
Coding Region ◽  
Typing Method ◽  
Genome Variability ◽  
Molecular Typing Method

To explore further the phylogenetic relationships between human enteroviruses and to develop new diagnostic approaches, we designed a pair of generic primers in order to study a 1452 bp genomic fragment (relative to the poliovirus Mahoney genome), including the 3′ end of the VP1-coding region, the 2A- and 2B-coding regions, and the 5′ moiety of the 2C-coding region. Fifty-nine of the 64 prototype strains and 45 field isolates of various origins, involving 21 serotypes and 6 strains untypeable by standard immunological techniques, were successfully amplified with these primers. By determining the nucleotide sequence of the genomic fragment encoding the C-terminal third of the VP1 capsid protein we developed a molecular typing method based on RT–PCR and sequencing. If field isolate sequences were compared to human enterovirus VP1 sequences available in databases, nucleotide identity score was, in each case, highest with the homotypic prototype (74.8 to 89.4%). Phylogenetic trees were generated from alignments of partial VP1 sequences with several phylogeny algorithms. In all cases, the new classification of enteroviruses into five identified species was confirmed and strains of the same serotype were always monophyletic. Analysis of the results confirmed that the 3′ third of the VP1-coding sequence contains serotype-specific information and can be used as the basis of an effective and rapid molecular typing method. Furthermore, the amplification of such a long genomic fragment, including non-structural regions, is straightforward and could be used to investigate genome variability and to identify recombination breakpoints or specific attributes of pathogenicity.

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