Home About us Contact | |||
Probe Hybridization (probe + hybridization)
Kinds of Probe Hybridization Selected AbstractsSarcoglycanopathies and the risk of undetected deletion alleles in diagnosis,,HUMAN MUTATION, Issue 1 2005Stefan J. White Abstract We have designed Multiplex Amplifiable Probe Hybridization (MAPH) probes for 28 exons of the sarcoglycan genes SGCA, SGCB, SGCG, and SGCD. The set was used to screen DNA from limb-girdle muscular dystrophy (LGMD) patients for the presence of pathogenic deletion or duplication mutations. An unexpected heterozygous deletion of SGCG exon 7 was detected in a patient from a consanguineous family in which a known c.525delT mutation segregates. The exon 7 deletion was inherited from the father, who was part of the consanguineous c.525delT branch of the family but who did not carry the c.525delT mutation. A similar, homozygous deletion had been identified in two unrelated LGMD patients from southern Italy. The deletion breakpoints were mapped, isolated, and sequenced, and were identical in all cases. Haplotype analysis showed the same alleles segregating with the mutation in all three patients, suggesting a common ancestor. Exonic deletions in sarcoglycanopathies appear to be rare events. However, we recommend screening for exonic deletions/duplications in patients where a mutation has not been identified in both alleles, as well as in seemingly homozygous cases where segregation of the mutations can not be confirmed in the parents. © 2005 Wiley-Liss, Inc. [source] Five-year maintenance follow-up of early-onset periodontitis patientsJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 6 2003Joanna J. Kamma Abstract Objectives: The purpose of this study was to evaluate the clinical and microbiological status of patients with early-onset or aggressive periodontitis (EOP) who had received supportive periodontal care (SPC) every 3,6 months for a period of 5 years, following active periodontal treatment. Material & Methods: The study population consisted of 25 individuals with early-onset periodontitis. Clinical examination and recordings of probing pocket depth (PPD) and clinical attachment level (CAL) were performed at baseline prior to treatment (T0), 3 months following the termination of active periodontal treatment (T1) and annually at the SPC appointments (T2,T3,T4,T5). Microbiological samples were obtained at the 5-year SPC (T5). Subgingival plaque samples for each individual were collected from one deep pocket (>5 mm), based on pretreatment measurements, randomly selected in each quadrant. The levels of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were determined using oligonucleotide probe hybridization. Results: During the 5-year period, the mean of SPC/patient was 12.7 sessions. A significant improvement was observed in PPD, CAL, gingival bleeding index and suppuration following treatment. However, between T1 and T5, 134 sites in 20 patients deteriorated with a CAL loss of,2 mm. Out of these 134 sites showing disease progression, microbial samples were randomly obtained in 13 sites (9.7%) from 8 patients. Among other factors, smoking and stress were found to have significant predictive value on the future attachment loss. P. gingivalis, T. denticola and total bacterial load were statistically significantly higher in patients who experienced disease progression during the 5-year maintenance period. Conclusions: For most EOP patients, regular SPC was effective in maintaining clinical and microbiological improvements attained after active periodontal therapy. However, a small percentage of sites was identified as progressive in 20 patients. Variables found to be related to periodontal progression were the presence of as well as the high bacterial counts of P. gingivalis, T. denticola and total bacterial load, number of acute episodes, number of teeth lost, smoking and stress. Zusammenfassung Erhaltungstherapie über fünf Jahre bei Patienten mit früh einsetzender Parodontitis (EOP) Ziele: Der Zweck dieser Studie war es, 5 Jahre nach aktiver Parodontalbehandlung den klinischen und mikrobiologischen Zustand von Patienten mit früh einsetzender oder aggressiver Parodontitis (EOP), bei welchen alle 3-6 Monate eine parodontale Erhaltungstherapie (SPC) erfolgte, zu evaluieren. Material & Methoden: Die Studienpopulation bestand aus 25 Individuen mit früh einsetzender Parodontitis. Die klinische Untersuchung und Aufzeichnung der Sondierungstiefe (PPD) sowie des klinischen Attachmentniveaus (CAL) erfolgten bei der Eingangsuntersuchung vor der Behandlung (T0), drei Monate nach Beendigung der aktiven Parodontalbehandlung (T1) und jährlich bei den SPC-Terminen (T2,T3,T4,T5). Die mikrobiologischen Proben wurden bei der 5-Jahres-SPC gewonnen (T5). Für jedes Individuum wurden die subgingivalen Plaqueproben in jedem Quadranten aus einer tiefen Tasche (>5mm) entnommen. Dies geschah randomisiert und auf der Grundlage der Messungen vor der Behandlung. Das Niveau von Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis und Treponema denticola wurden unter Verwendung der Hybridisierung mit Oligonukleotid-Sonden bestimmt. Ergebnisse: Während der 5-jährigen Periode betrug die mittlere Anzahl der SPC-Sitzungen pro Patient 12,7. Nach der Behandlung wurden bei PPD, CAL, Gingiva-Blutungs-Index und der Pusentleerung signifikante Verbesserungen beobachtet. Jedoch haben sich zwischen T1 und T5 bei 20 Patienten 134 Taschen mit einem CAL-Verlust von=2mm verschlechtert. Bei 8 Patienten wurden aus diesen 134 Taschen, mit Progression der Erkrankung, von 13 Taschen (9,7%) randomisiert mikrobiologische Proben entnommen. Innerhalb anderer Faktoren wurde bei Rauchen und Stress ein signifikanter Vorhersagewert für zukünftigen Attachmentverlust vorgefunden. Bei den Patienten, die in der 5-jährigen Erhaltungsperiode eine Progression der Erkrankung erfuhren lagen P. gingivalis, T. denticola und die bakterielle Gesamtbelastung höher. Schlussfolgerungen: Für die meisten EOP-Patienten die regelmäßig an der parodontalen Erhaltungstherapie teilnahmen war diese hinsichtlich der Aufrechterhaltung der nach der aktiven Parodontaltherapie erzielten klinischen und mikrobiologischen Verbesserungen erfolgreich. Jedoch wurde bei 20 Patienten ein geringer Prozentsatz von Taschen als fortschreitend identifiziert. Die Variablen, von denen gefunden wurde, dass sie eine Beziehung zur Progression haben waren: sowohl Vorhandensein von P. gingivalis, T. denticola als auch hohe Bakterienzahl von P. gingivalis, T. denticola und die bakterielle Gesamtbelastung, Anzahl der akuten Episoden, Anzahl verlorener Zähne, Rauchen und Stress. Résumé Suivi en maintenance sur 5 ans de patients atteints de parodontites d'apparition précoce. Objectifs: Cette étude se propose d'évaluer l'état clinique et microbiologique de patients atteints de parodontites d'apparition précoce ou agressive (EOP) qui furent suivis en maintenance (SPC) tous les 3-6 mois pendant une période de 5 ans après un traitement parodontal actif. Matériel & Méthodes: La population étudiée consistait en 25 individus atteints de parodontites d'apparition précoce. L'examen clinique et l'enregistrement des profondeurs de poche (PPD) et du niveau d'attache (CAL) furent réalisés avant le traitement (T0), 3 mois après la fin du traitement actif (T1) et chaque année aux rendez vous de maintenance (T2,T3,T4,T5). Des échantillons microbiologiques furent prélevés lors de la maintenance à 5 ans (T5). La plaque sous-gingivale de chaque patient fut prélevée d'une poche profonde (>5mm), sur la base des examens initiaux, choisis au hasard dans chaque quadrant. Les niveaux d' Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis et Treponema denticola furent déterminés par hybridation par sonde d'oligonucleotides. Résultats: pendant la période d'examination de 5 ans, la moyenne des SPC par patient fut de 12.7 sessions. Une amélioration significative fut observée pour PPD, CAL, l'indice de saignement gingival et la suppuration suite au traitement. Cependant, entre T1 et T5, 134 sites chez 20 patients connurent une détérioration avec une perte d'attache de 2 mm. De ces 134 sites qui présentaient une progression de la maladie, des échantillons microbiologiques furent obtenus aléatoirement dans 13 sites (9.7%) chez 8 patients. Parmi d'autres facteurs, le tabagisme et le stress furent reconnus comme ayant une significative valeur prédictive pour de futures pertes d'attache. P. gingivalis, T. denticola et la charge bactérienne totale étaient de façon statistiquement significatif plus importants chez les patients chez qui la maladie progressait au cours des 5 ans de maintenance. Conclusions: pour la plupart des patients atteints d' EOP, des soins parodontaux de soutien réguliers sont efficaces pour maintenir les améliorations cliniques et microbiologiques obtenus par le traitement actif. Cependant, un petit pourcentage de sites progressait chez 20 patients. Les variables en ralation avec cette progression étaient la présence et aussi un comptage important de P. gingivalis, T. denticola et la charge bactérienne totale, le nombre d'épisodes aigus le nombre de dents perdues le tabagisme et le stress. [source] Human leukocyte antigen-DRB1*1101 correlates with less severe hepatitis in Taiwanese male carriers of hepatitis B virus,JOURNAL OF MEDICAL VIROLOGY, Issue 4 2009Yi-Wen Huang Abstract Human leukocyte antigen (HLA) class II molecules are associated with host immune responses against hepatitis B virus infection. Male gender is the apparent host factor when someone encounters with the severity of hepatitis. The aim of this study was to investigate the association of the most polymorphic HLA class II allele, human leukocyte antigen,DRB1, with the severity of hepatitis in male carriers of hepatitis B virus. In this prospective cohort study, a total of 204 carriers of hepatitis B virus (131 men and 73 women) who have been followed-up for more than 1 year at the outpatient clinic of a university hospital were collected consecutively. Fifty carriers of hepatitis B virus (group I) with alanine aminotransferase <2× upper limit of normal (mean follow-up 83.6 months) were compared with 154 chronic hepatitis B patients (group II) with alanine aminotransferase ,2× upper limit of normal (mean follow-up 81.3 months). Alleles of HLA-DRB1 were typed by the polymerase chain reaction,sequence specific oligonucleotide probe hybridization and genotypes of hepatitis B virus by melting curve analysis. HLA-DRB1*1101 was found in 18% of group I versus 8% of group II in male carriers (OR 0.23, P,=,0.020, after adjustment for age) and 4% versus 9.4% in female carriers (P,=,0.094). In male carriers harboring DRB1*1101, the distribution of hepatitis B viral genotype was comparable between the two groups. HLA-DRB1*1101 correlates with less severe hepatitis in Taiwanese male carriers of hepatitis B virus. J. Med. Virol. 81:588,593, 2009 © 2009 Wiley-Liss, Inc. [source] HLA phenotypes and outcomes of hepatitis B virus infection in Taiwan,JOURNAL OF MEDICAL VIROLOGY, Issue 1 2004Ya-Fang Wu Abstract The relationship of HLA phenotype and outcome of hepatitis B virus (HBV) infection was studied in two ethnic groups of Taiwan: Han Chinese and Taiwanese Aborigines. In Han Chinese, the study groups consisted of 98 persons who tested both hepatitis B surface antigen (HBsAg) and anti-HBs negative (Uninfected Group), 324 persons who tested HBsAg negative and both anti-HBs and anti-HBc positive (Recovered Group), and 98 patients who tested HBsAg positive for at least 6 months (Chronically Infected Group). In Taiwanese Aborigines, the study groups consisted of 34 persons in Uninfected Group, 229 persons in the Recovered Group, and 138 patients in the Chronically Infected Group. All subjects were tested for HLA (A, B, DRB1) phenotypes by sequence-specific oligonucleotide probe hybridization (SSOPH). HLA-DR*0406 was significantly more frequent in the Recovered Group, compared with the Chronically Infected Group (P,<,0.001) in Han Chinese. There was a significant excess of HLA-B*4001 (P,=,0.045) in the Recovered Group, compared with the Chronically Infected Group in Taiwanese Aborigines. The observation that different HLA phenotypes associated with recovery from HBV infection in different racial groups implies that various HLA molecules could present different HBV epitopes to induce effective immune responses. J. Med. Virol. 72:17,25, 2004. © 2004 Wiley-Liss, Inc. [source] Epidemiology of Candidemia in a Turkish tertiary care hospital,APMIS, Issue 9 2006MUSTAFA BAKIR In order to determine the local epidemiology of candidemia, Candida strains isolated between 1994 and 2000 were identified to species level; antifungal resistance patterns and DNA fingerprints were analyzed. Identification of Candida strains (n: 140) was performed with germ tube test and carbohydrate assimilation reactions. Minimal inhibitory concentrations were determined using a commercial test for 5-flucytosine and the broth macrodilution method according to NCCLS for fluconazole and amphotericin B. Molecular relatedness was determined by restriction endonuclease analysis of genomic DNA followed by probe hybridization. C. albicans (37.2%), C. parapsilosis (32.2%), and C. tropicalis (12.2%) comprised 114 (81.4%) of 140 isolates. Susceptibility tests did not reveal resistance to amphotericin B in any of the Candida isolates. Fluconazole resistance was detected in one isolate of C. krusei, and 5-flucytosine resistance in two C. tropicalis isolates and one C. albicans isolate. Significantly higher frequency of clusters with identical strains in C. parapsilosis and C. tropicalis was detected compared to C. albicans. Pediatric wards are particularly important in the nosocomial transmission of non- albicans candida species. [source] HLA DPB1*0201 allele is negatively associated with immunoglobulin E responsiveness specific for house dust mite allergens in TaiwanCLINICAL & EXPERIMENTAL ALLERGY, Issue 4 2000Background House dust mite (HDM) Dermatophagoides pteronyssinus is the most important source of indoor allergens that cause allergic diseases in Taiwan. We prepared purified HDM allergens (Der p 1, Der p 2 and Der p 5) to detect allergen-specific immunoglobulin (Ig) E responsiveness among a large number of test subjects. The robust genetic typing system for HLA class II genes also facilitated the study on association of HLA and allergic response toward HDM. Objective This study intended to investigate the association between HLA class II alleles and the IgE responsiveness to the major allergens from HDM, D. pteronyssinus. Methods Two hundred and forty-eight subjects were selected for HLA association study. Plasma HDM allergen (Der p 1, Der p 2, Der p 5) -specific IgE and Der p 2-specific IgG antibodies were detected by ELISA, while HLA class II -DRB1, -DQA1, -DQB1, -DPB1 genetic polymorphism was determined by polymerase chain reaction/sequence-specific oligonucleotide probe hybridization (PCR/SSOPH). Statistical comparison of the allelic distribution of each HLA class II genes among the individuals with/without HDM allergen-specific IgE and IgG antibodies were performed. Results There was no significant association between HLA DRB1, DQB1, DQA1 alleles and HDM-specific IgE responsiveness noted. Only DRB1*0803 and the linked DQA1*0103 alleles showed positive association with Der p 5-specific IgE responsiveness. However, we found that HLA-DPB1*1301 predisposed subjects to IgE responsiveness to HDM Der p 5. HLA DPB1*0501 was weakly associated with the IgE responsiveness to HDM Der p 1 and Der p 5. There was a strong negative association between the HLA-DPB1*0201 allele with IgE responsiveness to Der p 1 (OR: 0.30, P , 0.0001, P , 0.0007, Pc , 0.010). Conclusion We clearly observed the association between HLA DPB1 alleles and specific IgE responsiveness to HDM major allergens. The molecular mechanism of HLA-DPB1*0201 involvement in protecting subjects from HDM-specific IgE responsiveness awaits further investigation. [source] Vero cytotoxin-producing Escherichia coli in a study of infectious intestinal disease in EnglandCLINICAL MICROBIOLOGY AND INFECTION, Issue 3 2002J. Evans An investigation of infectious intestinal disease in England included examination of feces for Vero cytotoxin-producing Escherichia coli (VTEC). Using DNA probe hybridization 27 VTEC strains were identified, 12 were from cases, and of these three belonged to serogroup O157. The remaining 15 strains were isolated from controls. The strains were confirmed biochemically as E. coli, they were serotyped and characterized according to their toxin production, the presence of sequences encoding intimin (eae) and enterohemolysin was determined and resistance to antimicrobial agents was determined. Six of the nine cases with non-O157 VTEC were less than 16 years old, only two of the 15 controls were under 16. Infection with more than one micro-organism was also considered. [source] |