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1.
Ocul Immunol Inflamm ; : 1-6, 2024 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-38324651

RESUMO

PURPOSE: To quantify chorioretinal microvascular damage and recovery post-treatment in patients with acute syphilitic posterior placoid chorioretinitis (ASPPC) using fractal dimension (FD). METHODS: Retrospective cohort study of patients with serologically confirmed syphilitic uveitis. We obtained optical coherence tomography angiography (OCTA) scans at baseline and follow-up after intravenous penicillin treatment and computed FD of the superficial capillary plexus (SCP), deep capillary plexus (DCP), and choriocapillaris (CC) using ImageJ. RESULTS: We enrolled seven patients with ASPPC (11 eyes), and 17 control subjects (34 eyes). Pre-treatment averages of FD-SCP, FD-DCP, and FD-CC were: 1.672 (±0.115), 1.638 (±0.097), and 1.72 (±0.137); post-treatment: 1.760 (±0.071), 1.764 (±0.043), and 1.898 (±0.047). After treatment FD-CC increased in all 11 eyes with an average of 0.163 (p = 0.003); FD-DCP increased in 10 (91%) eyes with an average of 0.126 (p = 0.003); and FD-SCP increased in seven (64%) eyes with an average of 0.089 (p = 0.059). Compared to the post-treatment FD values in the syphilitic group, controls had similar FD-SCP (p = 0.266), FD-DCP (p = 0.078), and FD-CC (p = 0.449). CONCLUSIONS: CC and DCP are mostly affected in ASPPC with minimal changes in the SCP. All vascular layers FD recovered after completing antibiotic treatment.

2.
JAC Antimicrob Resist ; 5(6): dlad118, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38021040

RESUMO

Introduction: Vancomycin-resistant Enterococcus faecium (VRE) meningitis accounts for only 0.3%-4.0% of bacterial meningitis cases and typically occurs following neurosurgical intervention. We describe a rare case of a patient without neurological devices in situ or a recent neurological procedure who developed VRE meningitis via haematogenous spread. Optimal treatment for VRE meningitis is unknown. Case presentation: A 67-year-old male with end-stage liver failure underwent liver transplantation complicated by VRE bacteraemia and subsequently developed VRE meningitis while on high-dose daptomycin therapy (12 mg/kg/day). Due to clinical and microbiological failure with daptomycin, he was switched to linezolid and symptoms resolved rapidly. He completed 2 weeks of linezolid, fully recovered, and continued to do well without complications at the 5 month follow-up. Conclusions: This case highlights the severity of VRE infections in solid organ transplant recipients and raises concerns about daptomycin penetration into the CNS. Linezolid could be considered the preferred treatment for VRE CNS infections rather than high-dose daptomycin.

3.
Monoclon Antib Immunodiagn Immunother ; 42(2): 65-67, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36787476

RESUMO

This study reports on hemodynamic changes observed during monoclonal antibody (mAb) administration for patients with severe acute respiratory distress syndrome-coronavirus-2. Findings from this study may have implications for patient safety. Hemodynamic data from 705 patients who received subcutaneous or intravenous mAb therapy during February 1, 2021-September 30, 2021 in clinics in Arkansas, USA were reviewed. Descriptive statistics and paired t-tests were used to assess blood pressure before and after treatment. Results showed 386 (54.7%) patients experienced a drop in systolic blood pressure (SBP) or diastolic blood pressure (DBP) >5 mmHg. The average drop in SBP was 9.2 mmHg for those patients. Two hundred and eighty-one (39.9%) patients experienced a drop in SBP of >10 mmHg with an average drop in SBP of 12.0 mmHg. The Emergency Use Authorization for mAb does not list hypotension as a contraindication for treatment. Our findings suggest mAb therapy should be administered in an environment where vitals are monitored.


Assuntos
Anticorpos Monoclonais , COVID-19 , Humanos , Pressão Sanguínea , Estudos Retrospectivos , SARS-CoV-2
4.
IDCases ; 31: e01645, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36579145

RESUMO

Objective: to describe a case of severe sepsis and complicated bacteremia caused by Arcanobacterium haemolyticum and review similar cases in the literature. Case summary: A 26-year-old gentleman with a history of epilepsy presented with symptoms of sore throat, productive cough, periumbilical abdominal pain, watery diarrhea, nausea and vomiting, subjective fevers along with progressive jaundice for seven days. The patient had acute fulminant liver failure, septic shock, and Multi-organ failure. He required vasopressors, underwent intubation, and had grown Arcanobacterium haemolyticum in the blood and Bronchoalveolar lavage samples. He developed a peritonsillar abscess and cavitary pneumonia and required chest tube drainage followed by thoracotomy for hemothorax. The patient improved on Ampicillin-Sulbactam treatment and was treated with a total antibiotic duration of 6 weeks. He fully improved on post-discharge follow-up. Discussion: Arcanobacterium haemolyticum is a Gram-positive (sometimes Gram variable), catalase-negative facultatively anaerobic, non-motile, non-spore-forming, and variably ß-hemolytic and is known to be a cause of pharyngitis and skin and soft tissue infections. Rarely A. Haemolyticum can be associated with severe systemic infections such as infective endocarditis, systemic abscesses, osteomyelitis, and septicemia. In previous literature reviews, the source of A. haemolyticum depended on the host, and pharyngeal and upper respiratory sources were likely to be associated with immunocompetent hosts. Conclusion: A. haemolyticum should be included in the differential diagnosis of bacterial pharyngitis complicated by severe systemic illness. Penicillins are the most commonly used antibiotics for treating A. haemolyticum bacteremia, and macrolides can be used for Penicillin's treatment failure.

5.
HIV Med ; 23(3): 268-273, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34725910

RESUMO

OBJECTIVES: A standardized non-occupational post-exposure prophylaxis (nPEP) programme was implemented to improve guideline compliance for treatment of post-sexual assault patients within an emergency department (ED). METHODS: A single-centre, retrospective, observational study of adult patients evaluated in the ED for sexual assault was performed following nPEP programme implementation. A comprehensive nPEP programme consisting of a standardized order set, real-time multidisciplinary consultation, on-site pharmacy and close post-discharge follow-up was implemented between July 2017 and June 2018. Laboratory, treatment, vaccination, prescription and follow-up data during the pre- (July 2016 to June 2017) and post-intervention (July 2018 to August 2019) periods were compared. RESULTS: Of the 147 post-sexual assault patients included in this study (59 pre-intervention, 88 post-intervention), 133 (90.5%) were eligible for nPEP. Patient demographics and rate of those eligible for nPEP were similar in both cohorts. Antiretroviral therapy (ART) was offered (72.2% vs. 100%; p < 0.005) and ultimately prescribed (51.9% vs. 86.1%; p < 0.005) more frequently following nPEP programme implementation. Patients were more likely to have appropriate screening for renal function, liver function, pregnancy, syphilis, hepatitis B, hepatitis C and HIV in the post-intervention period (all p < 0.005). Hepatitis B vaccination was more commonly administered post-intervention (8.5% vs. 22.7%; p < 0.024). In-person 28-day follow-up was rare in both pre- (3.5%) and post-intervention (11.3%) cohorts (p = 0.278). CONCLUSIONS: Implementation of a comprehensive nPEP programme resulted in improved guideline compliance with more frequent and appropriate ART administration. Recommended screening laboratories and hepatitis B vaccinations were more commonly performed, but in-person follow-up remained low. The nPEP programmes should be implemented to standardize efforts that decrease the risk of HIV transmission.


Assuntos
Infecções por HIV , Delitos Sexuais , Adulto , Assistência ao Convalescente , Serviço Hospitalar de Emergência , Infecções por HIV/tratamento farmacológico , Infecções por HIV/prevenção & controle , Humanos , Alta do Paciente , Profilaxia Pós-Exposição/métodos , Estudos Retrospectivos
6.
Clin Infect Dis ; 75(2): 269-277, 2022 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-34718456

RESUMO

BACKGROUND: Bloodstream infections (BSIs) are a leading cause of morbidity and mortality. The Improving Outcomes and Antimicrobial Stewardship study seeks to evaluate the impact of the Accelerate PhenoTest BC Kit (AXDX) on antimicrobial use and clinical outcomes in BSIs. METHODS: This multicenter, quasiexperimental study compared clinical and antimicrobial stewardship metrics, prior to and after implementation of AXDX, to evaluate the impact this technology has on patients with BSIs. Laboratory and clinical data from hospitalized patients with BSIs (excluding contaminants) were compared between 2 arms, 1 that underwent testing on AXDX (post-AXDX) and 1 that underwent alternative organism identification and susceptibility testing (pre-AXDX). The primary outcomes were time to optimal therapy (TTOT) and 30-day mortality. RESULTS: A total of 854 patients with BSIs (435 pre-AXDX, 419 post-AXDX) were included. Median TTOT was 17.2 hours shorter in the post-AXDX arm (23.7 hours) compared with the pre-AXDX arm (40.9 hours; P<.0001). Compared with pre-AXDX, median time to first antimicrobial modification (24.2 vs 13.9 hours; P<.0001) and first antimicrobial deescalation (36.0 vs 27.2 hours; P=.0004) were shorter in the post-AXDX arm. Mortality (8.7% pre-AXDX vs 6.0% post-AXDX), length of stay (7.0 pre-AXDX vs 6.5 days post-AXDX), and adverse drug events were not significantly different between arms. Length of stay was shorter in the post-AXDX arm (5.4 vs 6.4 days; P=.03) among patients with gram-negative bacteremia. CONCLUSIONS: For BSIs, use of AXDX was associated with significant decreases in TTOT, first antimicrobial modification, and time to antimicrobial deescalation.


Assuntos
Anti-Infecciosos , Gestão de Antimicrobianos , Bacteriemia , Infecções por Bactérias Gram-Negativas , Antibacterianos/uso terapêutico , Anti-Infecciosos/uso terapêutico , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Humanos
7.
Infect Control Hosp Epidemiol ; 43(3): 291-297, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-33896442

RESUMO

OBJECTIVE: To determine patient-specific risk factors and clinical outcomes associated with contaminated blood cultures. DESIGN: A single-center, retrospective case-control risk factor and clinical outcome analysis performed on inpatients with blood cultures collected in the emergency department, 2014-2018. Patients with contaminated blood cultures (cases) were compared to patients with negative blood cultures (controls). SETTING: A 509-bed tertiary-care university hospital. METHODS: Risk factors independently associated with blood-culture contamination were determined using multivariable logistic regression. The impacts of contamination on clinical outcomes were assessed using linear regression, logistic regression, and generalized linear model with γ log link. RESULTS: Of 13,782 blood cultures, 1,504 (10.9%) true positives were excluded, leaving 1,012 (7.3%) cases and 11,266 (81.7%) controls. The following factors were independently associated with blood-culture contamination: increasing age (adjusted odds ratio [aOR], 1.01; 95% confidence interval [CI], 1.01-1.01), black race (aOR, 1.32; 95% CI, 1.15-1.51), increased body mass index (BMI; aOR, 1.01; 95% CI, 1.00-1.02), chronic obstructive pulmonary disease (aOR, 1.16; 95% CI, 1.02-1.33), paralysis (aOR 1.64; 95% CI, 1.26-2.14) and sepsis plus shock (aOR, 1.26; 95% CI, 1.07-1.49). After controlling for age, race, BMI, and sepsis, blood-culture contamination increased length of stay (LOS; ß = 1.24 ± 0.24; P < .0001), length of antibiotic treatment (LOT; ß = 1.01 ± 0.20; P < .001), hospital charges (ß = 0.22 ± 0.03; P < .0001), acute kidney injury (AKI; aOR, 1.60; 95% CI, 1.40-1.83), echocardiogram orders (aOR, 1.51; 95% CI, 1.30-1.75) and in-hospital mortality (aOR, 1.69; 95% CI, 1.31-2.16). CONCLUSIONS: These unique risk factors identify high-risk individuals for blood-culture contamination. After controlling for confounders, contamination significantly increased LOS, LOT, hospital charges, AKI, echocardiograms, and in-hospital mortality.


Assuntos
Injúria Renal Aguda , Sepse , Choque Séptico , Hemocultura , Feminino , Humanos , Masculino , Estudos Retrospectivos , Fatores de Risco
8.
J Antimicrob Chemother ; 76(9): 2453-2463, 2021 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-34021752

RESUMO

BACKGROUND: Data from the Improving Outcomes and Antibiotic Stewardship for Patients with Bloodstream Infections: Accelerate PhenoTest™ BC Kit (AXDX) Registry Study were analysed to determine the impact of rapid organism identification and antimicrobial susceptibility testing (AST) for Gram-positive bacteraemia. PATIENTS AND METHODS: This multicentre, quasi-experimental study evaluated clinical and antimicrobial stewardship metrics following the implementation of AXDX. Data from hospitalized patients with bacteraemia were compared between groups, one that underwent testing on AXDX (post-AXDX) and one that underwent traditional identification and AST (pre-AXDX). An analysis of patients with Gram-positive bacteraemia was performed. The primary outcome was time to optimal therapy (TTOT). Secondary outcomes included time to first antibiotic modification (overall and Gram-positive), duration of unnecessary MRSA coverage, incidence of adverse events, length of stay and mortality. RESULTS: A total of 219 (109 pre-AXDX, 110 post-AXDX) patients with Gram-positive bacteraemia were included. Median TTOT was 36.3 h (IQR, 16.9-56.7) in the pre-AXDX group and 20.4 h (IQR, 7.5-36.7) in the post-AXDX group (P = 0.01). Compared with pre-AXDX, median time to first antibiotic modification (29.1 versus 15.9 h; P = 0.002), time to first Gram-positive antibiotic modification (33.2 versus 17.2 h; P = 0.003) and median duration of unnecessary MRSA coverage (58.4 versus 29.7 h; P = 0.04) were reduced post-AXDX. A trend towards decreased acute kidney injury (24% versus 13%; P = 0.06) was observed in the post-AXDX group. Groups did not differ in other secondary outcomes. CONCLUSIONS: Implementation of AXDX testing for patients with Gram-positive bacteraemia shortened the TTOT and reduced unnecessary antibiotic exposure due to faster antibiotic modifications.


Assuntos
Gestão de Antimicrobianos , Bacteriemia , Antibacterianos/uso terapêutico , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Humanos
9.
Clin Infect Dis ; 73(11): e4616-e4626, 2021 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-32463864

RESUMO

BACKGROUND: Accelerate Pheno blood culture detection system (AXDX) provides rapid identification and antimicrobial susceptibility testing results. Limited data exist regarding its clinical impact. Other rapid platforms coupled with antimicrobial stewardship program (ASP) real-time notification (RTN) have shown improved length of stay (LOS) in bacteremia. METHODS: A single-center, quasi-experimental study of bacteremic inpatients before and after AXDX implementation was conducted comparing clinical outcomes from 1 historical and 2 intervention cohorts (AXDX and AXDX + RTN). RESULTS: Of 830 bacteremic episodes, 188 of 245 (77%) historical and 308 (155 AXDX, 153 AXDX + RTN) of 585 (65%) intervention episodes were included. Median LOS was shorter with AXDX (6.3 days) and AXDX + RTN (6.7 days) compared to historical (8.1 days) (P = .001). In the AXDX and AXDX + RTN cohorts, achievement of optimal therapy (AOT) was more frequent (93.6% and 95.4%, respectively) and median time to optimal therapy (TTOT) was faster (1.3 days and 1.4 days, respectively) compared to historical (84.6%, P ≤ .001 and 2.4 days, P ≤ .001, respectively). Median antimicrobial days of therapy (DOT) was shorter in both intervention arms compared to historical (6 days each vs 7 days; P = .011). Median LOS benefit during intervention was most pronounced in coagulase-negative Staphylococcus bacteremia (P = .003). CONCLUSIONS: LOS, AOT, TTOT, and total DOT significantly improved after AXDX implementation. Addition of RTN did not show further improvement over AXDX and an already active ASP. These results suggest that AXDX can be integrated into healthcare systems with an active ASP even without the resources to include RTN.


Assuntos
Anti-Infecciosos , Bacteriemia , Antibacterianos/uso terapêutico , Anti-Infecciosos/uso terapêutico , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Hemocultura , Humanos , Staphylococcus
12.
Clin Infect Dis ; 67(9): 1356-1363, 2018 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-29668884

RESUMO

Background: Daptomycin-associated myopathy has been identified in 2%-14% of patients, and rhabdomyolysis is a known adverse effect. Although risk factors for daptomycin-associated myopathy are poorly defined, creatine phosphokinase (CPK) monitoring and temporary discontinuation of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, or "statins," has been recommended. Methods: We conducted a single-center, retrospective, matched case-control risk factor analysis in adult and pediatric patients from 2004 to 2015. Patients in whom myopathy (defined as CPK values above the upper limit of normal) developed during daptomycin treatment were matched 1:1 to no-myopathy controls with at least the same duration of therapy. Risk factors independently associated with myopathy were determined using multivariable conditional logistic regression. Secondary analysis was performed in patients with rhabdomyolysis, defined as CPK values ≥10 times the upper limit of normal. Results: Of 3042 patients reviewed, 128 (4.2%) were identified as having daptomycin-associated myopathy, 25 (0.8%) of whom had rhabdomyolysis; 121 (95%) of the 128 were adults, and the mean duration of therapy before CPK elevation was 16.7 days (range, 1-58 days). In multivariate analysis, deep abscess treatment (odds ratio, 2.80; P = .03), antihistamine coadministration (3.50; P = .03), and statin coadministration (2.60; P = .03) were independent risk factors for myopathy. Obesity (odds ratio, 3.28; P = .03) and statin coadministration (4.67; P = .03) were found to be independent risk factors for rhabdomyolysis, and older age was associated with reduced risk (0.97; P = .05). Conclusions: Statin coadministration with daptomycin was independently associated with myopathy and rhabdomyolysis. This is the first study to provide strong evidence supporting this association. During coadministration, we recommend twice-weekly CPK monitoring and consideration of withholding statins.


Assuntos
Antibacterianos/efeitos adversos , Daptomicina/efeitos adversos , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Doenças Musculares/induzido quimicamente , Rabdomiólise/induzido quimicamente , Adulto , Idoso , Antibacterianos/administração & dosagem , Estudos de Casos e Controles , Creatina Quinase/sangue , Daptomicina/administração & dosagem , Quimioterapia Combinada/efeitos adversos , Registros Eletrônicos de Saúde , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Tennessee
13.
Infect Dis Clin North Am ; 30(4): 1053-1070, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27816139

RESUMO

Health care-acquired viral respiratory infections are common and cause increased patient morbidity and mortality. Respiratory syncytial virus and influenza virus are frequently transmitted in the hospital setting. Studies report decreased nosocomial transmission when aggressive infection control measures are implemented with more success using a multicomponent approach. Influenza vaccination of health care personnel has been shown to further decrease rates of transmission, thus mandatory vaccination is becoming more common. This article focuses on the epidemiology, transmission, and control of health care-associated respiratory viral infections.


Assuntos
Infecção Hospitalar , Infecções Respiratórias , Viroses , Infecções por Coronavirus , Humanos , Influenza Humana
14.
J Ark Med Soc ; 108(10): 208-10, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22479977

RESUMO

Brown recluse spiders are predominantly found in south central United States. Their bites usually cause mild self-limiting reactions, although localized tissue necrosis and rare systemic, potentially fatal, envenomations are known to occur. Herein, we report an atypical presentation of a brown recluse bite in a 20 year old female who was admitted to the intensive care unit due to angioedema and cellulitis. We photographically document the bite site for twenty-four hours following envenomation. She received glucocorticoids, antihistamines, antibiotics and dapsone while hospitalized and was subsequently discharged with complete resolution of symptoms without the development of tissue necrosis or scarring.


Assuntos
Anafilaxia/etiologia , Mordeduras e Picadas/complicações , Lábio/lesões , Diester Fosfórico Hidrolases/efeitos adversos , Venenos de Aranha/efeitos adversos , Aranhas , Anafilaxia/patologia , Animais , Arkansas , Mordeduras e Picadas/patologia , Feminino , Humanos , Lábio/patologia , Adulto Jovem
15.
PLoS One ; 5(11): e15098, 2010 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-21152047

RESUMO

BACKGROUND: We describe the epidemiology of hospitalized RSV infections for all age groups from population-based surveillance in two rural provinces in Thailand. METHODS: From September 1, 2003 through December 31, 2007, we enrolled hospitalized patients with acute lower respiratory tract illness, who had a chest radiograph ordered by the physician, from all hospitals in SaKaeo and Nakhom Phanom Provinces. We tested nasopharyngeal specimens for RSV with reverse transcriptase polymerase chain reaction (RT-PCR) assays and paired-sera from a subset of patients with IgG enzyme immunoassay. Rates were adjusted for enrollment. RESULTS: Among 11,097 enrolled patients, 987 (8.9%) had RSV infection. Rates of hospitalized RSV infection overall (and radiographically-confirmed pneumonia) were highest among children aged<1 year: 1,067/100,000 (534/100,000 radiographically-confirmed pneumonia) and 1-4 year: 403/100,000 (222/100,000), but low among enrolled adults aged≥65 years: 42/100,000. Age<1 year (adjusted odds ratio [aOR]=13.2, 95% confidence interval [CI] 7.7, 22.5) and 1-4 year (aOR=8.3, 95% CI 5.0, 13.9) were independent predictors of hospitalized RSV infection. CONCLUSIONS: The incidence of hospitalized RSV lower respiratory tract illness among children<5 years was high in rural Thailand. Efforts to prevent RSV infection could substantially reduce the pneumonia burden in children aged<5 years.


Assuntos
Infecções por Vírus Respiratório Sincicial/epidemiologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/epidemiologia , População Rural/estatística & dados numéricos , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Pré-Escolar , Hospitalização/estatística & dados numéricos , Humanos , Incidência , Lactente , Recém-Nascido , Pessoa de Meia-Idade , Pneumonia/complicações , Pneumonia/epidemiologia , Vigilância da População , RNA Viral/genética , Infecções por Vírus Respiratório Sincicial/complicações , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/genética , Infecções Respiratórias/complicações , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tailândia/epidemiologia , Adulto Jovem
16.
Emerg Infect Dis ; 14(3): 465-7, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18325263

RESUMO

Acanthamoeba polyphaga mimivirus (APM), a virus of free-living amebae, has reportedly caused human respiratory disease. Using 2 newly developed real-time PCR assays, we screened 496 respiratory specimens from 9 pneumonia-patient populations for APM. This virus was not detected in any specimen, which suggests it is not a common respiratory pathogen.


Assuntos
Acanthamoeba/virologia , Vírus de DNA/isolamento & purificação , Pneumonia/virologia , Animais , DNA Viral , Humanos , Reação em Cadeia da Polimerase
17.
J Clin Microbiol ; 46(2): 533-9, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18057136

RESUMO

Human rhinoviruses (HRVs) are important contributors to respiratory disease, but their healthcare burden remains unclear, primarily because of the lack of sensitive, accurate, and convenient means of determining their causal role. To address this, we developed and clinically validated the sensitivity and specificity of a real-time reverse transcription-PCR (RT-PCR) assay targeting the viral 5' noncoding region defined by sequences obtained from all 100 currently recognized HRV prototype strains and 85 recently circulating field isolates. The assay successfully amplified all HRVs tested and could reproducibly detect 50 HRV RNA transcript copies, with a dynamic range of over 7 logs. In contrast, a quantified RNA transcript of human enterovirus 68 (HEV68) that showed the greatest sequence homology to the HRV primers and probe set was not detected below a concentration of 5 x 10(5) copies per reaction. Nucleic acid extracts of 111 coded respiratory specimens that were culture positive for HRV or HEV were tested with the HRV real-time RT-PCR assay and by two independent laboratories that used different in-house HRV/HEV RT-PCR assays. Eighty-seven HRV-culture-positive specimens were correctly identified by the real-time RT-PCR assay, and 4 of the 24 HEV-positive samples were positive for HRV. HRV-specific sequences subsequently were identified in these four specimens, suggesting HRV/HEV coinfection in these patients. The assay was successfully applied in an investigation of a coincidental outbreak of HRV respiratory illness among laboratory staff.


Assuntos
Infecções por Picornaviridae/diagnóstico , Infecções por Picornaviridae/virologia , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Rhinovirus/classificação , Rhinovirus/isolamento & purificação , Regiões 5' não Traduzidas/genética , Surtos de Doenças , Enterovirus/genética , Humanos , Dados de Sequência Molecular , Infecções por Picornaviridae/epidemiologia , RNA Viral/genética , Infecções Respiratórias/epidemiologia , Rhinovirus/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA
18.
J Infect Dis ; 196(9): 1321-8, 2007 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-17922396

RESUMO

BACKGROUND: We sought to determine whether infections with human coronaviruses (HCoVs) 229E, OC43, HKU1, and NL63 are associated with pneumonia and to define the epidemiology of HCoV infection in rural Thailand. METHODS: We developed a real-time reverse-transcription polymerase chain reaction (RT-PCR) assay panel for the recognized HCoV types and compared HCoV infections in patients hospitalized with pneumonia, outpatients with influenza-like illness, and asymptomatic control patients between September 2003 and August 2005. RESULTS: During study year 1, 43 (5.9%) of 734 patients with pneumonia had HCoV infections; 72.1% of the infections were with OC43. During study year 2, when control patients were available, 21 (1.8%) of 1156 patients with pneumonia, 12 (2.3%) of 513 outpatients, and 6 (2.1%) of 281 control patients had HCoV infections. Compared with infection in control patients, infection with any HCoV type or with all types combined was not associated with pneumonia (adjusted odds ratio for all HCoV types, 0.67 [95% confidence interval, 0.26-1.75]; P=.40). HCoV infections were detected throughout both study years; 93.6% of OC43 infections in the first year occurred from January through March. CONCLUSIONS: HCoV infections were infrequently detected in rural Thailand by use of sensitive real-time RT-PCR assays. We found no association between HCoV infection and illness. However, we noted year-to-year variation in the prevalence of HCoV strains, which likely influenced our results.


Assuntos
Infecções por Coronavirus/epidemiologia , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Infecções por Coronavirus/virologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Pneumonia Viral/epidemiologia , População Rural , Sensibilidade e Especificidade , Tailândia/epidemiologia
19.
J Clin Microbiol ; 45(7): 2105-9, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17507510

RESUMO

Clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, high-throughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. We evaluated two multiplex reverse transcription-PCR (RT-PCR) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens. The NGEN respiratory virus analyte-specific assay (Nanogen, San Diego, CA) detects influenza A virus (Flu-A) and Flu-B, parainfluenza virus 1 (PIV-1), PIV-2, and PIV-3, and respiratory syncytial virus (RSV), while the ResPlex II assay (Genaco Biomedical Products, Inc., Huntsville, AL) detects Flu-A, Flu-B, PIV-1, PIV-2, PIV-3, PIV-4, RSV, human metapneumovirus (hMPV), rhinoviruses (RhVs), enteroviruses (EnVs), and severe acute respiratory syndrome (SARS) coronavirus (CoV). A total of 360 frozen respiratory specimens collected for a full year were tested, and results were compared to those obtained with a combined reference standard of cell culture and monoplex real-time TaqMan RT-PCR assays. NGEN and ResPlex II gave comparable sensitivities for Flu-A (82.8 to 86.2%), Flu-B (90.0 to 100.0%), PIV-1 (87.5 to 93.8%), PIV-3 (66.7 to 72.2%), and RSV (63.3 to 73.3%); both assays achieved excellent specificities (99.1 to 100.0%) for these five common viruses. The ResPlex II assay detected hMPV in 13 (3.6%) specimens, with a sensitivity of 80.0% and specificity of 99.7%. The ResPlex II assay also differentiated RSV-A and RSV-B and gave positive results for RhV and EnV in 31 (8.6%) and 19 (5.3%) specimens, respectively. PIV-2, PIV-4, and SARS CoV were not detected in the specimens tested. The two systems can process 80 (NGEN) and 96 (ResPlex II) tests per run, with a hands-on time of approximately 60 min and test turnaround times of 6 h (ResPlex II) and 9 h (NGEN). Multiple-panel testing detected an additional unsuspected 9 (3.4%) PIV-1 and 10 (3.7%) PIV-3 infections. While test sensitivities for RSV and PIV-3 need improvement, both the NGEN and ResPlex II assays provide user-friendly and high-throughput tools for simultaneous detection and identification of a panel of common respiratory viral pathogens in a single test format. The multiplex approach enhances diagnosis through detection of respiratory viral etiologic agents in cases in which the presence of the agent was not suspected and a test was not ordered by the clinicians.


Assuntos
Infecções por Vírus de RNA/virologia , Vírus de RNA/isolamento & purificação , Doenças Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Viroses/virologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Vírus de RNA/classificação
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