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1.
BMC Health Serv Res ; 23(1): 220, 2023 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-36882832

RESUMO

BACKGROUND: The UK National Health Service (NHS) is ideally placed to undertake research. The UK Government recently launched its vision of research within the NHS to improve research culture and activity amongst its staff. Currently, little is known about the research interest, capacity and culture of staff in one Health Board in South East Scotland and how their attitudes to research may have changed as a result of the SARS-CoV-2 pandemic. METHODS: We used the validated Research Capacity and Culture tool in an online survey of staff working in one Health Board in South East Scotland to explore attitudes to research at the organisation, team and individual level together with involvement in, barriers to and motivators to engage in research. Questions included changes in attitude to research as a result of the pandemic. Staff were identified by professional group: nurses/midwives, medical/dental, allied health professionals (AHP), other therapeutic and administrative roles. Median scores and interquartile ranges were reported and differences between groups assessed using the Chi-square and Kruskal-Wallis tests with P < 0.05 accepted as statistical significance. Free-text entries were analysed using content analysis. RESULTS: Replies were received from 503/9145 potential respondents (5.5% response), of these 278 (3.0% response) completed all sections of the questionnaire. Differences between groups were noted in the proportions of those with research as part of their role (P = 0.012) and in being research-active (P < 0.001). Respondents reported high scores for promoting evidence-based practice and for finding and critically reviewing literature. Low scores were returned for preparing reports and securing grants. Overall, medical and other therapeutic staff reported higher levels of practical skills compared with other groups. Principal barriers to research were pressure of clinical work and lack of time, backfill and funds. 171/503 (34%) had changed their attitude to research as a result of the pandemic with 92% of 205 respondents more likely to volunteer for a study themselves. CONCLUSION: We found a positive change in attitude to research arising from the SARS-CoV-2 pandemic. Research engagement may increase after addressing the barriers cited. The present results provide a baseline against which future initiatives introduced to increase research capability and capacity may be assessed.


Assuntos
COVID-19 , Pandemias , Humanos , Estudos Transversais , COVID-19/epidemiologia , SARS-CoV-2 , Medicina Estatal , Escócia/epidemiologia
2.
J Obstet Gynaecol Res ; 47(2): 689-697, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33274569

RESUMO

OBJECTIVE: To determine the value of uterine CD138+ cells, as a marker of chronic endometritis, in predicting subsequent reproductive outcome in women with history of recurrent pregnancy loss. DESIGN: A prospective longitudinal study. SETTING: Tertiary specialized clinic. PATIENTS: Women with history of recurrent pregnancy loss or implantation failure over a 12-months follow-up period. INTERVENTION: We quantified the CD138+ cells/high powered field (hpf) using immunohistochemistry and image analysis of endometrial biopsies obtained during the secretory stage post ovulation. MAIN OUTCOME MEASURES: Live birth and subsequent pregnancy loss. We calculated the receiver operator curve for predicting subsequent pregnancy loss and reported using relative risk (RR) and 95% confidence intervals (CI). RESULTS: We enrolled 344 women of whom 88 became pregnant (88/344, 25.5%). Half of them had a subsequent live birth (47/88, 53%) and the rest lost their pregnancy (41/88, 46%). The median CD138+ score was significantly lower in the live birth group (P < 0.005) and women with a CD138+ score ≥ 16/hpf had a higher risk of subsequent miscarriage (RR 10.0, 95% CI 2.78-36.02). CD138+ cells count showed a good prediction for subsequent pregnancy loss in high-risk women with an area under the curve of 0.75 (95% CI 0.59-0.82, P = 0.01). A cut-off value of 4-6 cells/hpf offered the best predictive accuracy with higher scores predicting worse reproductive outcome. Our findings are limited by the small event rate and the sample size of our cohort. CONCLUSION: Quantifying CD138+ cells by immunohistochemistry in women with a history of recurrent pregnancy loss is helpful to diagnose chronic endometritis and predict subsequent reproductive outcome.


Assuntos
Aborto Habitual , Endometrite , Estudos de Coortes , Endometrite/diagnóstico , Endometrite/epidemiologia , Endométrio , Feminino , Humanos , Estudos Longitudinais , Gravidez , Estudos Prospectivos
3.
Clin Kidney J ; 10(6): 822-829, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29225812

RESUMO

BACKGROUND: Exercise improves physical capacity in patients with end-stage renal disease on haemodialysis (HD), but few patients engage in it. Whole-body vibration exercise (WBVE) is a novel protocol that has been shown to benefit frail elderly patients' rehabilitation. We assessed the utility of WBVE before HD sessions and tested methods to inform the design of a randomized controlled trial (RCT). METHODS: Physical condition and quality of life were assessed at enrolment and repeated 2 weeks later in a pilot study of 49 patients undergoing regular HD. All patients then undertook 8 weeks of WBVE, thrice weekly for 3 min, after which the assessments were repeated and results compared (paired t-tests). Further assessments were made after a 4-week layoff. Patients completed a post-study questionnaire about their experiences of using WBVE. The reproducibility of WBVE and effects on measures of functionality, muscle strength, indirect exercise capacity, nutritional status, bone health and quality of life were recorded to undertake a power calculation for an RCT. RESULTS: Of 49 patients enrolled, 25 completed all assessments. The dropout rate was high at 49%, but overall, WBVE was an acceptable form of exercise. Functionality as assessed by the 60-s sit-to-stand test (STS-60) improved significantly by 11% (P = 0.002). Some quality of life domains also improved significantly. All improvements were maintained 4 weeks after discontinuing WBVE. CONCLUSIONS: WBVE was acceptable, safe, easily incorporated into the routine of HD and was associated with useful improvements in physical function sufficient to justify a RCT.

4.
Aust N Z J Psychiatry ; 48(11): 977-1008, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25351912

RESUMO

OBJECTIVES: This clinical practice guideline for treatment of DSM-5 feeding and eating disorders was conducted as part of the Royal Australian and New Zealand College of Psychiatrists (RANZCP) Clinical Practice Guidelines (CPG) Project 2013-2014. METHODS: The CPG was developed in accordance with best practice according to the National Health and Medical Research Council of Australia. Literature of evidence for treatments of anorexia nervosa (AN), bulimia nervosa (BN), binge eating disorder (BED), other specified and unspecified eating disorders and avoidant restrictive food intake disorder (ARFID) was sourced from the previous RANZCP CPG reviews (dated to 2009) and updated with a systematic review (dated 2008-2013). A multidisciplinary working group wrote the draft CPG, which then underwent expert, community and stakeholder consultation, during which process additional evidence was identified. RESULTS: In AN the CPG recommends treatment as an outpatient or day patient in most instances (i.e. in the least restrictive environment), with hospital admission for those at risk of medical and/or psychological compromise. A multi-axial and collaborative approach is recommended, including consideration of nutritional, medical and psychological aspects, the use of family based therapies in younger people and specialist therapist-led manualised based psychological therapies in all age groups and that include longer-term follow-up. A harm minimisation approach is recommended in chronic AN. In BN and BED the CPG recommends an individual psychological therapy for which the best evidence is for therapist-led cognitive behavioural therapy (CBT). There is also a role for CBT adapted for internet delivery, or CBT in a non-specialist guided self-help form. Medications that may be helpful either as an adjunctive or alternative treatment option include an antidepressant, topiramate, or orlistat (the last for people with comorbid obesity). No specific treatment is recommended for ARFID as there are no trials to guide practice. CONCLUSIONS: Specific evidence based psychological and pharmacological treatments are recommended for most eating disorders but more trials are needed for specific therapies in AN, and research is urgently needed for all aspects of ARFID assessment and management. EXPERT REVIEWERS: Associate Professor Susan Byrne, Dr Angelica Claudino, Dr Anthea Fursland, Associate Professor Jennifer Gaudiani, Dr Susan Hart, Ms Gabriella Heruc, Associate Professor Michael Kohn, Dr Rick Kausman, Dr Sarah Maguire, Ms Peta Marks, Professor Janet Treasure and Mr Andrew Wallis.


Assuntos
Antidepressivos/uso terapêutico , Terapia Cognitivo-Comportamental/métodos , Transtornos da Alimentação e da Ingestão de Alimentos/terapia , Fármacos Antiobesidade/uso terapêutico , Austrália , Doença Crônica , Transtornos da Alimentação e da Ingestão de Alimentos/complicações , Transtornos da Alimentação e da Ingestão de Alimentos/psicologia , Frutose/análogos & derivados , Frutose/uso terapêutico , Redução do Dano , Humanos , Lactonas/uso terapêutico , Nova Zelândia , Obesidade/complicações , Obesidade/tratamento farmacológico , Orlistate , Psiquiatria , Sociedades Médicas , Topiramato
5.
F1000Res ; 2: 31, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24358863

RESUMO

BACKGROUND: Arthroscopic shoulder surgery is increasingly performed as a day case procedure. Optimal post-operative pain relief remains a challenge due to considerable variations in the level of pain experienced between individuals. Our aim was to examine whether the preoperative electrical pain threshold was a strong predictor of elevated postoperative pain levels following arthroscopic subacromial decompression (ASD) surgery. METHODS: Forty consenting patients with American Society of Anesthesiologists (ASA) grade 1-2 presenting for elective ASD surgery were recruited. Patients' electrical pain thresholds were measured preoperatively using a PainMatcher® (Cefar Medical AB, Lund, Sweden) device. Following surgery under general anaesthesia, the maximum pain experienced at rest and movement was recorded using a visual analogue scale until the end of postoperative day four. RESULTS: In univariate analyses (t-test), the postoperative pain experienced (Area Under Curve) was significantly greater in patients with a low pain threshold as compared with a high pain threshold at both rest (mean 12.5, S.E. 1.7 v mean 6.5, S.E.1.2. P=0.008) and on movement (mean 18.7, S.E. 1.5 v mean 14.1, S.E.1.4. P=0.031). In multivariate analyses, adjusting for additional extra analgesia, the pain experienced postoperatively was significantly greater in the low pain threshold group both at rest (mean difference 4.9, 95% CI 1.5 to 8.4, P=0.007) and on movement (mean difference 4.1, 95%CI 0.03 to 8.2, P=0.049). CONCLUSIONS: Preoperative pain threshold can predict postoperative pain level following ASD of the shoulder. TRIAL REGISTRATION: Clinicaltrials.gov identifier: NCT01351363 LEVEL OF EVIDENCE: II.

6.
Ophthalmic Surg Lasers Imaging ; 42(3): 229-33, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21563746

RESUMO

BACKGROUND AND OBJECTIVE: To determine posterior capsule opacification (PCO) rate and identify contributory factors in a series of patients undergoing combined 23-gauge transconjunctival phacovitrectomy. PATIENTS AND METHODS: Retrospective data on 221 consecutive patients operated on by a single senior surgeon using a standard technique were collected and subjected to univariate analysis with chi-square tests and multivariate analysis with logistic regression. RESULTS: PCO developed in 46 of 221 cases (20.8%). Major risk factors were: rhegmatogenous retinal detachment (relative risk = 3.3, P = .002), axial length greater than 24.5 mm (relative risk = 2.0, P = .093), intraoperative/postoperative complications (relative risk = 3.3, P = .04), C2F6 tamponade (versus SF6/air) (relative risk = 2.8, P = .01), and postoperative posturing (relative risk = 4.1, P < .001). Multiple logistic regression analysis showed the latter two to be most important. CONCLUSION: PCO rate following 23-gauge phacovitrectomy is lower than most rates reported for 20-gauge phacovitrectomy. Using shorter-acting gas tamponade and avoiding postoperative posturing may help lower PCO rates.


Assuntos
Opacificação da Cápsula/etiologia , Microcirurgia/efeitos adversos , Facoemulsificação/efeitos adversos , Cápsula Posterior do Cristalino/patologia , Vitrectomia/efeitos adversos , Adulto , Idoso , Idoso de 80 Anos ou mais , Opacificação da Cápsula/diagnóstico , Catarata/complicações , Túnica Conjuntiva , Feminino , Fluorocarbonos/administração & dosagem , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Doenças Retinianas/complicações , Doenças Retinianas/cirurgia , Estudos Retrospectivos , Fatores de Risco , Hexafluoreto de Enxofre/administração & dosagem , Adulto Jovem
7.
Sex Transm Infect ; 87(5): 433-8, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21429898

RESUMO

OBJECTIVE: To understand the influence of voluntary counselling and testing (VCT) and an HIV-negative result on subsequent sexual behaviour; and to identify the specific felt needs of those testing negative. DESIGN: Serial interviews, 6 months apart, with people who had tested negative, and three one-off focus group discussions with counsellors. Interviews were transcribed verbatim and analysed thematically. Paired transcripts were analysed longitudinally. PARTICIPANTS: Purposively sampled from first attenders for VCT who had tested negative. SETTING: Four health-facility affiliated VCT centres in two mining towns in the Copperbelt Province of Zambia. RESULTS: Participants in the 42 initial and 31 follow-up interviews understood VCT as 'testing for HIV'. Most participants reported having adopted safer sexual behaviour months before attending VCT. They had already recognised personal susceptibility to HIV infection and reported engaging both emotionally and cognitively with the issues of testing. Ultimately attendance at testing was to know their status in order to regain control of their lives. Unmet post-test support needs were common and included additional information, supportive networks, life-skills training and access to recreational activities. CONCLUSIONS: In this population who tested HIV negative the majority said they had made life changes months before attending VCT. Counselling served to consolidate pre-test decisions about risk behaviour. Those testing HIV negative are underserved compared with those testing positive. We recommend that counselling for HIV should be de-linked from testing to encourage earlier attendance for counselling and that clients testing negative should receive more post-test support to help them remain HIV negative.


Assuntos
Aconselhamento , Infecções por HIV/diagnóstico , Soronegatividade para HIV , Comportamento Sexual , Adulto , Atitude Frente a Saúde , Preservativos/estatística & dados numéricos , Feminino , Infecções por HIV/psicologia , Humanos , Masculino , Estado Civil , Pessoa de Meia-Idade , Mineração , Aceitação pelo Paciente de Cuidados de Saúde , Sexo sem Proteção , Programas Voluntários , Adulto Jovem , Zâmbia
8.
Br J Gen Pract ; 60(581): e449-58, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21144189

RESUMO

BACKGROUND: Advance care planning is being promoted as a central component of end-of-life policies in many developed countries, but there is concern that professionals find its implementation challenging. AIM: To assess the feasibility of implementing advance care planning in UK primary care. DESIGN OF STUDY: Mixed methods evaluation of a pilot educational intervention. SETTING: Four general practices in south-east Scotland. METHOD: Interviews with 20 GPs and eight community nurses before and after a practice-based workshop; this was followed by telephone interviews with nine other GPs with a special interest in palliative care from across the UK. RESULTS: End-of-life care planning for patients typically starts as an urgent response to clear evidence of a short prognosis, and aims to achieve a 'good death'. Findings suggest that there were multiple barriers to earlier planning: prognostic uncertainty; limited collaboration with secondary care; a desire to maintain hope; and resistance to any kind of 'tick-box' approach. Following the workshop, participants' knowledge and skills were enhanced but there was little evidence of more proactive planning. GPs from other parts of the UK described confusion over terminology and were concerned about the difficulties of implementing inflexible, policy-driven care. CONCLUSION: A clear divide was found between UK policy directives and delivery of end-of-life care in the community that educational interventions targeting primary care professionals are unlikely to address. Advance care planning has the potential to promote autonomy and shared decision making about end-of-life care, but this will require a significant shift in attitudes.


Assuntos
Planejamento Antecipado de Cuidados/organização & administração , Medicina Geral/educação , Assistência Terminal/organização & administração , Atitude do Pessoal de Saúde , Estudos de Viabilidade , Humanos , Assistência Terminal/psicologia
9.
Fam Pract ; 26(4): 269-74, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19502575

RESUMO

BACKGROUND: Our current knowledge of the epidemiology of infertility is limited and outdated. Health care provision for infertility in the UK attracts public interest because of restrictions on access to services. OBJECTIVE: To describe the incidence, prevalence, referral patterns and outcomes of infertile couples, presenting in general practice in UK. METHODS: A population-based retrospective observational outcome study of infertile couples from general practices in Northumberland, Tyne and Wear, UK (population 1 043 513). Outcome data at 1 year were collected on all couples who presented to their GP between the 1st January 2005 and 30th June 2006 with a fertility problem. RESULTS: Thirty-four per cent of general practices in the study area contributed data (population 404 263). The incidence of infertility was 0.9 couples per 1000 general population. The average age of women was 31 years, and the average time attempting conception was 18 months. Treatment end points for half of all couples were in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Over half of the couples in the study were not eligible for National Health Service (NHS) fertility treatment on social criteria. At 12 months, 27% of all couples in the study achieved a pregnancy spontaneously and a further 9% with treatment. CONCLUSIONS: Infertile women present to their GP later in life compared with 20 years ago, and after a shorter period of infertility. Half of the couples required treatment with IVF or ICSI. Adopting the British Fertility Society recommendation of allowing couples, where one or both partners has a child in a previous relationship, will result in an additional 26% of infertile couples becoming eligible for NHS fertility treatment.


Assuntos
Infertilidade Feminina/epidemiologia , Infertilidade Masculina/epidemiologia , Adulto , Fatores Etários , Feminino , Humanos , Incidência , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/terapia , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/terapia , Masculino , Gravidez , Prevalência , Atenção Primária à Saúde , Ensaios Clínicos Controlados Aleatórios como Assunto , Técnicas de Reprodução Assistida , Estudos Retrospectivos , Medicina Estatal , Fatores de Tempo , Resultado do Tratamento , Reino Unido/epidemiologia , Adulto Jovem
10.
Med Decis Making ; 27(4): 438-47, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17641140

RESUMO

BACKGROUND: Stroke risk in nonvalvular atrial fibrillation can be reduced by warfarin or aspirin; the choice of therapy requires the assessment of risks and benefits. The authors compared methods of risk assessment and their implications for risk communication and treatment. METHODS: Stroke risk was compared in 193 patients with atrial fibrillation using the Framingham equation; an atrial fibrillation-specific Framingham equation; the Congestive heart failure, Hypertension, Age, Diabetes and Stroke (CHADS(2)) score; the Stroke Prevention and Atrial Fibrillation (SPAF) scheme; and the Scottish Intercollegiate Guidelines Network (SIGN) guidelines. Treatment guidance from SIGN, a simple prediction rule, and a decision analytical approach was compared. In the latter, patients were classified as risk too low to benefit from warfarin if the risk of cerebral bleeding on warfarin approximated to, or exceeded, thromboembolic stroke risk reduction. RESULTS: Framingham equations gave lower stroke risks overall than SIGN or SPAF. CHADS(2) was intermediate. Using SIGN, warfarin would be given to all 103 patients without a history of stroke/transient ischemic attack and for whom warfarin was not contraindicated but only to 73 patients using the simple prediction rule and 48 patients using the decision analysis. CONCLUSION: Community-based cohorts give lower stroke risk estimates than CHADS(2); both give lower estimates than schemes from control groups from randomized controlled trials. Using community-derived risks would lead to fewer patients being treated with warfarin than guidance derived from randomized controlled trial controls, which may lead to many low-risk patients being treated with high-risk therapy. This raises the debate about appropriate sources of data for risk assessment to support risk communication and effective clinical decisions.


Assuntos
Anticoagulantes/uso terapêutico , Fibrilação Atrial/tratamento farmacológico , Tomada de Decisões , Acidente Vascular Cerebral/prevenção & controle , Idoso , Idoso de 80 Anos ou mais , Fibrilação Atrial/complicações , Comorbidade , Medicina Baseada em Evidências , Feminino , Humanos , Masculino , Guias de Prática Clínica como Assunto , Medição de Risco , Fatores de Risco , Fatores Sexuais , Acidente Vascular Cerebral/etiologia , Varfarina
13.
Hum Fertil (Camb) ; 9(1): 47-51, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16581721

RESUMO

Infertility affects one in seven couples in the United Kingdom. The National Institute for Clinical Excellence (NICE) issued guidance on the management of the infertile couple in February 2004, which included the statement "for the assessment of tubal damage, women not known to have co-morbidities (pelvic inflammatory disease, endometriosis or previous ectopic pregnancy) should be offered hysterosalpingography (HSG)". We made HSG available to six general practices in Newcastle upon Tyne as an open access investigation. Our aim was to evaluate the uptake of open access HSG, speed of access to specialist services and the quality of the information recorded in the referral letter. Using hospital clinical records we tracked the outcome of all infertile couples from the six pilot practices over a nine-month period. Of the 39 referrals identified, 10 women were eligible for open access HSG, of which six HSGs were organized by GPs. Couples who had open access HSG reached a diagnosis and management plan four weeks earlier than those who were referred directly (mean difference 4.0 weeks, 95% confidence interval (CI) -8.8 to 0.4 weeks). The information recorded in the referral letter was generally poor. However, all referrals made via the open access HSG service had the prerequisite tests done. Open access HSG allowed prompter access to specialist services with more complete information passed on in the referral letter. Open access HSG was used in 15% of all infertile couples and 60% of those who fitted the criteria for its use. Open access HSG together with semen analysis and endocrine blood tests may allow GPs to manage the initial stages of the infertile couple and make a diagnosis.


Assuntos
Histerossalpingografia , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/terapia , Atenção Primária à Saúde , Adulto , Feminino , Hormônios/sangue , Humanos , Masculino , Projetos Piloto , Sêmen/citologia , Reino Unido
14.
J Clin Densitom ; 7(2): 134-42, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15181257

RESUMO

We assessed the utility of os calcis (OC) bone mineral density (BMD) measurements to identify men with low BMD at the lumbar spine (LS) and femoral neck (FN). BMD was measured by dual X-ray absorptiometry (DXA). Receiver operator characteristics (ROC) analysis was applied to determine the risk of osteoporosis at the lumbar spine or femoral neck. [A total of 230 men with an average age of 59 yr were studied.] The most common reasons for referral were fracture (47%) and steroid use (46%). Twenty-six percent were osteoporotic at the LS, 21% at the FN, and 15% at the OC. Optimal classification with respect to osteoporotic measurements at the LS or FN was obtained at an OC T-score of -1.9 (BMD = 0.45 g/cm2). Osteoporosis was only weakly related to a simple cumulative risk factor score, but was strongly related to a T-score OC categorized into quartiles. Regression analysis of BMD on the major risk factors alone explained only 17% of the variance in BMD at the LS and 5% at the FN. The combination of the T-score at the OC, age, and weight provided the best model. BMD OC is superior to risk factors alone in the clinical evaluation and selection of men referred for axial densitometry.


Assuntos
Densidade Óssea , Colo do Fêmur/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Funções Verossimilhança , Masculino , Pessoa de Meia-Idade , Prevalência , Curva ROC , Sensibilidade e Especificidade
16.
Br J Gen Pract ; 53(494): 714-5, 2003 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15103880

RESUMO

Domiciliary oxygen is expensive and is frequently used outside the prescribing guidelines, which include the need for blood oxygen measures, a hospital-based facility. Ongoing prescriptions are generally provided by general practitioners (GPs). A survey in the north-east of England found that the origin of the initial prescription was often obscure and that there was no record of the responsible clinician or of structured follow-up for the majority of patients. Many patients received oxygen outside the prescribing guidelines. There is a need for better organised, conjoint follow-up of patients on domiciliary oxygen.


Assuntos
Serviços de Assistência Domiciliar/organização & administração , Oxigenoterapia/normas , Idoso , Baixo Débito Cardíaco/terapia , Inglaterra , Medicina de Família e Comunidade/normas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oxigenoterapia/economia , Guias de Prática Clínica como Assunto , Padrões de Prática Médica/economia , Padrões de Prática Médica/normas , Doença Pulmonar Obstrutiva Crônica/terapia , Doença Cardiopulmonar/terapia
17.
Anal Biochem ; 307(1): 47-53, 2002 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-12137778

RESUMO

We report the development of a qualitative fluorescent multiplex homogeneous assay designed for the detection of the two most common hemochromatosis mutations using dual-labeled fluorescent probes. The assay is able to detect four allelic variants in a single closed tube using a single thermocycling protocol. The procedure combines the great sensitivity of the polymerase chain reaction, the specificity provided by allele-specific oligonucleotide hybridization using the 5(') nuclease assay format, and the higher throughput of a multicolor fluorescence detection procedure. Genomic DNA was prepared from whole blood specimens using standard procedures. Following DNA sample preparation, two regions of the hemochromatosis gene (HFE) including the H63D and C282Y mutations were coamplified and detected in real-time by four different fluorescently labeled allele-specific oligonucleotide probes. Assay specificity was demonstrated by a blind methods comparison study that included 37 DNA samples from individuals with a known HFE genotype. Results from the study showed that the multicolor multiplex HFE assay unambiguously classified all possible genotypes for the HFE gene C282Y and H63D mutations(1). This technique will be useful for research and molecular diagnostic laboratories and can be easily adapted for the detection of other single nucleotide polymorphisms.


Assuntos
Hemocromatose/genética , Antígenos de Histocompatibilidade Classe I/genética , Proteínas de Membrana/genética , Mutação/genética , Alelos , Análise Mutacional de DNA , Sondas de DNA/química , Sondas de DNA/genética , Corantes Fluorescentes , Genótipo , Hemocromatose/sangue , Proteína da Hemocromatose , Antígenos de Histocompatibilidade Classe I/sangue , Humanos , Proteínas de Membrana/sangue , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
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