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Semantic features of an enterprise interface terminology for SNOMED RT.

OBJECTIVE: To evaluate the utility of SNOMED RT in support of a natural language interface for encoding of clinical assessments. METHOD: Using a random sample of clinical terms from the UNMC Lexicon, I mapped the terminology into canonical data entries using SNOMED RT. Working from the source term language, I evaluated lexical mapping to the SNOMED term set, and the function of the SNOMED RT semantic network in support of a language-based clinical coding interface. RESULTS: Ambiguity in the source terms was low at 0.3%. Lexical (language-based) mapping could account for only 48.8% of meaning from the source terms. The RT semantic network accounted for 39.5% of meaning, and supplementing the lexical map this led to 80.2% capture of source content. Error rates in the segment of RT which I reviewed were low at 0.6%. 97.6% of source content could be accurately captured in SNOMED RT. CONCLUSION: SNOMED RT supported an accurate and reliable representation of clinical assessment data in this sample. The semantic network of RT substantially enhanced the encoding of concepts relative to lexical mapping. However these data suggest that natural language encoding with SNOMED RT in an enterprise environment is unlikely at this time.

Natural Language Processing↗

Efficacy of ebrotidine and ranitidine combined with amoxicillin and metronidazole in the eradication of Helicobacter pylori in patients with duodenal ulcer.

This double-blind, randomized, phase III clinical trial was carried out in two parallel groups to assess the efficacy of ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene) amino]-4-thiazolyl]methyl]thio]ethyl]amino] methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) 400 mg and ranitidine 300 mg given in single evening dose, combined with amoxicillin 750 mg and metronidazole 500 mg three times daily for 14 days, in the eradication of Helicobacter pylori in patients with duodenal ulcer. Thirty patients were included, divided into two groups of 15, to whom one of the study therapies was administered based on a randomization code. Clinical and endoscopic controls were performed 4, 6 and 8 weeks after the onset of the treatment. No differences were seen between the two treatment groups with regard to demographic parameters and clinical histories. They were both perfectly homogeneous. There were no differences between the eradication of both therapies in both the antrum and gastric body samples (over 80% eradication), allowing the results to be classified as satisfactory. Moreover, perfect control was achieved through the study of clinical symptoms, which even disappeared in some cases. There were no differences in the healing rate of the duodenal ulcer after four weeks, 86.7% being achieved for both groups.

Adult↗

Direct and uninterrupted RNA amplification of enteroviruses with colorimetric microwell detection.

BACKGROUND: Enteroviruses (EV) cause a broad spectrum of human diseases, of which aseptic meningitis is among the most common and most clinically vexing. While the clinical symptoms of meningitis caused by bacteria, fungi and viruses are similar, the diagnosis, therapy and outcome of disease caused by these agents vary greatly. In order to appropriately manage meningitis patients, rapid and reliable diagnosis of EV meningitis impacts significantly on patient management. OBJECTIVE: To develop a direct and uninterrupted RNA amplification of enteroviruses using rTth DNA polymerase. STUDY DESIGN: Purified coxsackievirus B6 RNA of various concentrations was amplified by rTth DNA polymerase-mediated amplification to determine analytic sensitivity. The specificity of the EV amplification was examined with a panel of nucleic acids from 36 EV serotypes, 15 non-EV pathogens and 10 coded clinical specimens of cerebrospinal fluid (CSF). RESULTS: All EV serotypes tested were detected successfully by this method at a sensitivity of 1 TCID(50) with the exception of echoviruses 1, 5, 22 and 23. Echovirus 5 was detected at 10 TCID(50), and echovirus 1 was detected at 100 TCID(50). Echoviruses 22 and 23 were not detectable at 100 TCID(50). Cross-reactivity of EV RT-PCR assay with 15 known non-EV meningitis pathogens has not been observed. Results of 10 CSF tested with this system correlated well with tissue culture. CONCLUSIONS: We have developed an EV amplification assay which has several important advantages over previously reported methods. This assay employs rTth DNA polymerase which possesses both reverse transcriptase and DNA polymerase activities, simplifying RNA reverse transcription and DNA amplification to an uninterrupted reaction. Additionally, potential carryover contamination and enhanced amplification specificity is provided by substituting dUTP for dTTP and adding uracil N-glycosylase (UNG) in the amplification reaction. Finally, the detection of amplified product is via a colorimetric, microwell format permitting the use of readily available instrumentation.

Journal Article↗

Transparent ICD and DRG coding using information technology: linking and associating information sources with the eXtensible Markup Language.

With the introduction of ICD-10 as the standard for diagnostics, it becomes necessary to develop an electronic representation of its complete content, inherent semantics, and coding rules. The authors' design relates to the current efforts by the CEN/TC 251 to establish a European standard for hierarchical classification systems in health care. The authors have developed an electronic representation of ICD-10 with the eXtensible Markup Language (XML) that facilitates integration into current information systems and coding software, taking different languages and versions into account. In this context, XML provides a complete processing framework of related technologies and standard tools that helps develop interoperable applications. XML provides semantic markup. It allows domain-specific definition of tags and hierarchical document structure. The idea of linking and thus combining information from different sources is a valuable feature of XML. In addition, XML topic maps are used to describe relationships between different sources, or "semantically associated" parts of these sources. The issue of achieving a standardized medical vocabulary becomes more and more important with the stepwise implementation of diagnostically related groups, for example. The aim of the authors' work is to provide a transparent and open infrastructure that can be used to support clinical coding and to develop further software applications. The authors are assuming that a comprehensive representation of the content, structure, inherent semantics, and layout of medical classification systems can be achieved through a document-oriented approach.

Diagnosis-Related Groups↗

Evaluation of electronic databases used to identify solid oral dosage forms.

The ability of electronic drug identification databases to identify solid oral dosage forms by their imprint codes was studied. The following seven commercially available electronic drug identification databases were selected to identify 500 solid oral dosage forms by their imprint codes: Clinical Pharmacology (Gold Standard Media, Tampa, FL), eFacts (Facts and Comparison, St. Louis, MO), Ident-A-Drug (Therapeutic Research, Stockton, CA), Identidex (Micromedex, Greenwood Village, CO), Clinical Reference Library (Lexi-Comp, Hudson, OH), Physicians' Desk Reference (PDR) Electronic Library (Medical Economics, Montvale, NJ), and RxList (RxList LLC, San Francisco, CA), Chi-square test was used to compare the percentages of medications identified by each of the seven electronic references. The ability of the databases to identify medication by specific characteristics, such as brand name versus generic, prescription versus nonprescription, commercially available for more than one year versus less than one year, colored versus white drug products, and controlled versus noncontrolled substances was evaluated. A logistic regression model was used to determine the probability of a drug product being identified by one of the electronic references based on these characteristics. All seven electronic databases combined identified 95.6% of the unknown medications by imprint code, color, shape, and scoring. Ident-A-Drug and Identidex identified the most drugs. The PDR Electronic Library and Facts and Comparisons Identified the least number of drugs. Solid oral dosage forms more likely to be identified were those that were on the market for more than a year, brand-name products, and prescription medications. Generic products on the market for less than a year and nonprescription products were particularly difficult to identify. A combination of electronic drug identification databases provides the best method of drug identification in an institutional setting.

Capsules↗

An association between length of stay and co-morbidity in chronic airflow limitation.

OBJECTIVE: To examine factors which impact on the length of stay and readmission for patients with chronic airflow limitation at a South Australian hospital from December 1996 until March 1998. DESIGN: Discharges from Flinders Medical Centre for patients aged > or = 18 years, where chronic airflow limitation was an active problem, and including a subset with a primary diagnosis of chronic airflow limitation, were identified, retrospectively, by the center's Clinical Coding Service from the hospital's in-patient separation database. SETTING: Flinders Medical Centre, Adelaide, South Australia. OUTCOME MEASURES: Length of stay; number of co-morbidities; readmission within 28 days. RESULTS: Five-hundred and twenty discharges (male:female, 258:262) with a primary diagnosis of chronic airflow limitation (ANDRG-3 177, chronic obstructive airways disease) were identified. Readmission within 28 days was related to the number of co-morbidities and to age. A relationship between length of stay and the number of co-morbidities was identified. A mean length of stay of 6.39 days was found for patients with less than five co-morbidities, 5.36 at their first admission to Flinders Medical Centre and 3.25 at their first admission to Flinders Medical Centre with no co-morbidities. These mean lengths of stay fall below overseas data previously published and are consistent with Kong's estimate of an ideal mean length of stay of 3.2 days when a clinical management guideline is used in low-risk chronic airflow limitation patients. CONCLUSIONS: Length of stay and readmission to hospital within 28 days of patients with a primary diagnosis of chronic airflow limitation is at least partly related to the number of co-morbidities and to age. The study has highlighted the difficulty of relying on changes to aggregate data as outcome measures for these patients.

Adult↗

Multidisciplinary team interventions for delirium in patients with chronic cognitive impairment.

BACKGROUND: Delirium is common in hospitalized elderly people. In the frail elderly, delirium may occur in 60% of those hospitalized. In the cognitively impaired, 45% have been shown to develop delirium and these patients have longer lengths of stay and a higher rate of complications which, amongst other things, together contribute to an increase in cost of care. The combination of being elderly and chronically cognitively impaired leads to a high risk of delirium with the associated increased risk of prolonged hospital stay, complications, and poor outcomes. The management of delirium has commonly been multifaceted - the primary emphasis has always been on the diagnosis and therapy of the precipitating factors, but as this may not be immediately resolved, symptomatic and supportive care may become of major importance. OBJECTIVES: The objective of this review is to assess the available evidence for the effectiveness, if any, of multidisciplinary team interventions in the coordinated care of patients with delirium superimposed on an underlying chronic cognitive impairment compared with the usual care of older cognitively impaired patients. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Cochrane Library, up to and including Issue 1, 1998) was searched using the terms 'delirium, controlled trial, cognitive'. MEDLINE, EMBASE and Psychlit (Ovid via Winspirs up to Feb 1998) were also searched with the same terms. Other sources including personal communications, ongoing trials, conference proceedings, handsearching and reference lists of published papers and books were all searched for relevant randomized controlled trials. The total yield from searching was 157 from which 8 (eight) were retained for consideration in the review. SELECTION CRITERIA: From the initial search yields, all randomised controlled trials involving the management of elderly patients with delirium were identified. A single reviewer (AMB) discarded irrelevant publications based on the title of the publication and its abstract. In the event that the article could possibly be relevant, it was retrieved for further assessment. All references were compiled in a list with a commentary on type of article, eg review, prospective study etc and this was independently considered by the second reviewer (RR) who agreed to review all randomised controlled studies reported on patients with delirium. Selection for possible inclusion in this review was then made on the basis of the participants reported as having chronic cognitive impairment, who then developed incident delirium and were randomly assigned to either coordinated multidisciplinary care or usual care. The outcomes of interest were length of stay in hospital, morbidity (including complications), patient distress & impact on care environment, mortality, discharge arrangements and follow-up including assessment of cognitive function at 6 months. Studies in which patients with chronic cognitive impairment or dementia, managed for incident delirium, according to ICD 9 criteria (see note) were considered eligible for inclusion in the review. Studies of risk factors and non-randomized studies were excluded. Note: this classification has been widely utilised throughout the English speaking medical literature over the past 20 years: ICD 10 is still being incorporated into clinical coding systems and has not been utilised in studies published in 1996. (ABSTRACT TRUNCATED)

Delirium↗

Severe chronic venous insufficiency: magnitude of the problem and consequences.

The aim of this study was to characterize patients requiring hospitalization for severe chronic venous insufficiency (CVI) at the local and national levels and to analyze factors related to primary amputation. An administrative database (Nationwide Inpatient Sample, 1988-2000) and a single institution (1992-2000) were reviewed using the International Classification of Diseases, 9th ed., Clinical Modification, codes for CVI, excluding phlegmasia and concomitant peripheral vascular occlusive disease codes. Demographics, clinical course, and outcomes were assessed. Descriptive, univariate, and multivariate statistical analyses were used; p < 0.05 was considered significant. Nationally, CVI occurred with a mean incidence of 92/100,000 admissions, of which 55% were women, having a mean age of 65 years and a median length of stay of 7 days. Mean hospital charges were $13,900 and did not change significantly over time. Acute deep vein thrombosis affected 1.3%, amputation was performed in 1.2%, and in-hospital mortality was 1.6% The local cohort included 67 patients with a mean age of 51 years; a majority were men (60%), and 85% were C6 (of Clinical-Etiologic-Anatomic-Pathophysiology [CEAP]). Patients averaged 23 clinic visits and a median of one hospitalization for CVI care over a 44-month follow-up. Twelve patients (18%) underwent a CVI-related amputation (one transmetatarsal amputation, nine below-knee amputations, and two above-knee amputations). They had fourfold more CVI-related hospitalizations, greater preoperative chronic narcotic use than nonamputee patients (85% vs. 58%), but less ongoing wound care needs (25% vs. 89%) (all p values < 0.05). However, no significant difference in long-term mortality, number of clinic visits, duration of symptoms, antibiotic courses, or prior venous-related surgeries was found. In those with amputation, ambulatory status was maintained in 75% at 15-month follow-up. The physiological and economic costs of severe CVI are significant and have not decreased over more than a decade. Amputation for CVI-related nonhealing wounds has a reasonable outcome. Future therapy must focus on prevention of CVI sequelae.

Aged↗

An evaluation of the relationship between specialist training in cardiology and implementation of evidence-based care of patients following acute myocardial infarction.

AIMS: Large clinical trials have provided evidence of prognostically beneficial treatment strategies for patients with acute myocardial infarction. However, the implementation of this evidence into routine clinical practice is suboptimal. We hypothesised that the speciality of the attending physician (cardiologist or not) would affect the use of evidence-based strategies. METHODS: Over a 3-month period (1st September to 30th November 1995), 3684 consecutive potential cases of acute myocardial infarction (AMI) in 20 adjacent hospitals in the Yorkshire Region were identified from coronary care registers, clinical coding and biochemistry records of cardiac enzyme assay requests. There were 2153 consecutive cases of AMI identified, of which 1643 patients were alive at discharge. We compared the admission use of aspirin and thrombolysis, and the use of aspirin, beta-blockers, angiotensin-converting enzyme (ACE) inhibitors and statins at discharge between cardiologists and other physicians. RESULTS: AMI patients under the care of cardiologists are more likely to receive aspirin and thrombolysis on the day of their event and to be prescribed aspirin, beta-blockers and statins on discharge. After correction for contraindications to their use, the above findings were broadly confirmed. DISCUSSION: Cardiologists are more likely than general physicians to use evidence-based treatment strategies recognised to improve AMI patient outcome. It is likely that this will translate into a reduction of mortality or other hard endpoints in patient outcomes.

Adrenergic beta-Antagonists↗

Gestational trophoblastic disease among Hispanic women: a 21-year hospital-based study.

OBJECTIVE: Hispanics are the fastest growing minority group in the United States. Few reports have described gestational trophoblastic disease (GTD) in this population. The purpose of this study was to determine the incidence of GTD at our public hospital which primarily serves the Hispanic population. METHODS: All women diagnosed with GTD (partial and complete hydatidiform mole, choriocarcinoma) between 1983 and 2004 were identified from the institutional tumor registry, surgical pathology reports and hospital ICD-9 codes. Clinical data were retrospectively extracted from medical records. The live birth denominator was tabulated over the same interval of time by retrieving labor and delivery statistics and sorting by race. RESULTS: GTD was diagnosed in 596 patients over a 21-year study interval encompassing 289,897 live births. The overall incidence of GTD was 2.06/1000 live births. Hispanic women had a higher incidence compared to Blacks (2.38 vs. 1.34; P < 0.001), but not Whites (2.00; P = 0.17). The 416 Hispanic women were diagnosed with GTD at an earlier gestational age in the latter part of this study (12.3 vs. 16.2 weeks; P < 0.001). Hispanics were more likely to have a partial hydatidiform mole compared to Blacks (29% vs. 13%; P < 0.001) and Whites (18%; P = 0.04). Choriocarcinomas occurred least commonly in Hispanic patients (1 per 35,000 live births). Teenage Hispanic women were the only ethnic age group with a higher risk of developing GTD (odds ratio = 1.6, 95% confidence interval: 1.1, 2.2). CONCLUSION: Hispanic women had the highest incidence of GTD in this hospital-based study, were diagnosed at an earlier gestational age in the last decade and more frequently were diagnosed with partial moles.

Adolescent↗

Factors associated with hospitalization costs for patients with community-acquired pneumonia.

BACKGROUND: Combined mortality rates for pneumonia and influenza suggest that the 2 conditions represent the sixth leading cause of death in the United States. The total cost of pneumonia, including indirect costs, was estimated to be approximately $23 billion per year in 1994. OBJECTIVE: The objective of this study was to assess variables that may be significantly associated with the cost of treating patients hospitalized with community-acquired pneumonia (CAP). We also assessed the impact of treatment guidelines for management of CAP (developed by managed care plans) on total costs. METHODS: Patients in 3 managed care plans who were hospitalized with a primary or secondary diagnosis of CAP in Maryland and Washington, DC, between January 1, 1997, and April 30, 1997, or between January 1, 1998, and April 30, 1998, were identified based on International Classification of Diseases, Ninth Revision codes. Clinical data were abstracted from patients' medical charts by nurses, and billing data were acquired from these plans. A retrospective data analysis was carried out using billing data from 3 managed care plans and clinical data from hospitals associated with the plans. A multivariate regression model was developed using the natural logarithm of cost as the dependent variable. Independent variables that were studied included severity of illness, days in the intensive care unit (ICU), triage per guidelines, drug therapy per guidelines, mortality, and managed care plan identifiers. RESULTS: The charts of 569 patients were assessed. The mean age of the study sample was 75.3 years. ICU days (P < 0.001), mortality, and drug therapy (both P < 0.01) per guidelines significantly affected costs. As expected, an increase in the number of ICU days led to an increase in costs. However, patients who received drug therapy recommended by the guidelines had significantly lower costs than patients not treated according to the guidelines (P = 0.001). CONCLUSIONS: The findings of this study suggest that guidelines for CAP management, such as those developed by managed care plans, may help reduce costs by minimizing unnecessary ICU admissions and appropriately managing patients with CAP.

Aged↗

Anabolic profiles in late-stage cancer patients responsive to hydrazine sulfate.

The anabolic profiles of 59 late-stage cancer patients responsive to hydrazine sulfate were examined; the drug had been given either as a sole agent or added to preexisting therapy to which the patients had become refractory. Most of the patients (79.7%) responded which Indicated Appetite Improvement (IAI), expressed by protocol-code, clinical evaluation and/or direct quantitation. In those patients receiving hydrazine sulfate alone the IAI was 86.1%; in those in whom hydrazine sulfate was added to pre-existing therapy the IAI was 69.6%. Of those cases expressed in direct quantitation the average weight gain for patients receiving hydrazine sulfate alone was 8.2 lbs, whereas the average weight gain for those with pre-existing therapy was 0.6 lbs (p = 0.01). The results suggest the use of hydrazine sulfate as a specific chemotherapy for cancer cachexia, and implicate ineffective concurrent or prior therapy as an apparent negative factor in the generation of anabolic response.

Appetite↗

Risk factors for trachoma in Mali.

OBJECTIVES: Prior to commencing a campaign to eliminate blinding trachoma in Mali, a national disease prevalence survey was conducted from March 1996 to June 1997. The prevalence of trachoma was estimated and potential risk factors were studied. METHODS: In each of Mali's seven regions (excluding the capital Bamako), a sample of 30 clusters was taken from the general population, in accordance with the principle of probability proportional to the size of the community. All children under 10 years of age were examined. The simplified clinical coding system proposed by the World Health Organization was used. The position of each village was established and subsequently related to the nearest meteorological station. Socioeconomic and environmental information was collected at both village and household level. The mother or caretaker of each child was questioned about availability and use of water for washing the child. At the time of examination, facial cleanliness and the presence of flies on the face were noted. RESULTS: A total of 15,187 children under 10 years of age were examined. The prevalence of active trachoma (follicular [TF] or intense trachoma [TI]) was 34.9% (95% CI : 32.3-37.6) and the prevalence of TI was 4.2% (95% CI : 3.5-5.0). Aridity/environmental dryness appears to be a risk factor influencing the current geographical distribution of trachoma. Small villages had considerably higher trachoma prevalence than their larger neighbours. The proximity of a medical centre and the existence of social organizations such as a women's association were associated with lower levels of trachoma. Crowded living conditions increased the risk. Using a monetary marker of wealth, we observed a linear inverse relation between wealth and trachoma prevalence. The presence of a dirty face was strongly associated with trachoma (odds ratio [OR] = 3.67) as was the presence of flies on the child's face (OR = 3.62). Trachoma prevalence increased with distance to a water source. Disease prevalence decreased with a higher frequency of both face washing and bathing. CONCLUSIONS: Of all the risk factors examined, facial cleanliness had the strongest association with the prevalence of trachoma. This was followed by the presence of flies on the child's face. Both face washing and bathing showed beneficial effects. Socioeconomic factors such as wealth were significantly explanatory. It is likely that hygiene education and fly control by environmental improvement could have a very significant impact on the prevalence of trachoma in Mali.

Baths↗

Impact of antisocial lifestyle on health.

BACKGROUND: Antisocial lifestyle includes a range of related behaviours which constitute threats to health. Links between health outcomes and particular behaviours have been investigated, but the overall impact on health is largely unknown. In part, this reflects lack of longitudinal studies designed to link offending and other antisocial behaviour, injury and illness. METHODS: Injury and illness data were collected prospectively in the longitudinal Cambridge Study of Delinquent Development (CSDD) for the age ranges 16-18 and 27-32. These data were translated into Read clinical codes, version 3.1 and categorized. Three hypotheses underpinned this investigation: that links between (i) convictions and injury and between (ii) childhood predictors of delinquency and injury at age 16-18 would be maintained at age 27-32; that (iii) antisocial behaviour at age 16-18 would be linked to injury at age 27-32; and that (iv) observed links between antisocial behaviour and decreased illness at age 16-18 would be reversed by age 27-32. RESULTS: Childhood predictors of teenage offending predicted injury and cardiovascular and psychological illness at age 27-32. Delinquency predicted road, home and self injury and psychological disorder, but was associated with less illness and hospital admission. Measures of antisocial behaviour at age 18 which most strongly predicted illness/injury at age 32 were drunk driving, fighting after drinking, sexual promiscuity and heavy smoking. Relationships at age 16-18 between heavy alcohol consumption and less infection and less organic illness remained at age 27-32. Links common to both age 16-18 and age 27-32 were those between convictions, heavy regular drinking, low job status and injury; and links between convictions and less organic illness. Principal differences included links between convictions and psychological disorders only found at age 27-32. CONCLUSIONS: Antisocial lifestyle was associated with injury and psychological disorders up to age 32 but also with less organic illness. Alcohol consumption at age 16-18 predicted fewer illnesses overall, and fewer infections 15 years later. These negative associations are important and deserve further study.

Adolescent↗

Highly active antiretroviral therapy and the incidence of HIV-1-associated nephropathy: a 12-year cohort study.

OBJECTIVE: to assess temporal changes in the incidence of human immunodeficiency virus-1-associated nephropathy (HIVAN), and the association with use of highly active antiretroviral therapy (HAART). METHODS: HIVAN incidence and risk factors were assessed in 3976 HIV-1-infected individuals followed in clinical cohort in Baltimore, Maryland, USA from 1989 to 2001. The incidence of HIVAN, defined by biopsy or a conservative uniformly applied clinical coding protocol, was expressed in terms of person-years, and Poisson regression was used for multivariate analysis. RESULTS: Ninety-four patients developed HIVAN over the course of the study for an incidence of 8.0 per 1000 person-years [95% confidence interval (CI), 6.5 to 9.8]. African American race and advanced immunosuppression were strongly associated with HIVAN risk. HIVAN incidence declined significantly in 1998-2001 compared with 1995-1997. Among patients with a prior diagnosis of AIDS, HIVAN incidence was 26.4, 14.4, and 6.8 per 1000 person-years in patients not receiving antiretroviral therapy, treated with nucleoside analogue therapy only, or treated with HAART, respectively (P < 0.001 for trend). In multivariate analysis, HIVAN risk was reduced 60% (95% CI, -30 to -80%) by use of HAART, and no patient developed HIVAN when HAART had been initiated prior to the development of AIDS. CONCLUSION: HAART was associated with a substantial reduction in HIVAN incidence. Additional follow-up will be needed to determine if renal damage in susceptible individuals is halted or merely slowed by HAART, particularly when control of viremia is incomplete or intermittent.

AIDS-Associated Nephropathy↗

Rapid serological technique for typing herpes simplex viruses.

A rapid technique is described which can accurately identify a herpes simplex virus (HSV) isolate as type 1 or type 2. Filter paper disks were used to immobilize viral antigens, which were then identified by means of an (125)I-labeled staphylococcal protein A immunoassay. The assay was performed in a specially designed 96-well filtration device which served as both an incubation chamber and a filter manifold. By using this system and cross-absorbed antisera to HSV types 1 and 2, 69 coded clinical isolates of HSV were correctly and unequivocally typed. HSV was also clearly distinguished from varicella-zoster virus and cytomegalovirus. This assay can be rapidly executed (less than 2 h) and yielded an objective endpoint; it required only minute quantities of typing sera and can be easily performed with the cells from a single infected roller tube culture. Thus, it can be used to type initial clinical isolates of HSV, yielding results within hours after the first appearance of cytopathic effects in the culture used for primary virus isolation. Moreover, it is particularly well suited to the simultaneous analysis of many specimens and is amenable to automation. These characteristics suggest that this (125)I-labeled staphylococcal protein A immunofiltration technique will be applicable to the rapid identification of other herpesviruses, as well as other clinical isolates.

Antigens, Viral↗

Enhanced enzyme-linked immunosorbent assay for detection of antibodies to virus-like particles of human papillomavirus.

Measurement of antibodies to human papillomavirus (HPV) is complicated by many factors. Although enzyme-linked immunosorbent assays (ELISAs) that use virus-like particles (VLPs) have proved useful, the assays have, in general, had moderate sensitivities and low signal-to-noise ratios. To enhance the performance of the assay, a systematic investigation was undertaken to examine key variables used in ELISAs for the detection of antibodies to VLPs of HPV. Incorporation of two vinyl polymers, polyvinyl alcohol (molecular weight, 50,000) (PVA-50) and polyvinylpyrrolidone (molecular weight, 360,000) (PVP-360), was found to increase the sensitivity as well as the specificity of the assay for the detection of antibodies to VLPs of HPV. In particular, the addition of PVA-50 to the blocking solution reduced the amount of nonspecific binding of antibodies to VLPs and the microplate surface, whereas the addition of PVP-360 increased the sensitivity of antibody detection. The new ELISA demonstrated increased sensitivity and specificity for the detection of cervical HPV type 16 infection compared to those of a prototype assay with coded clinical serum samples from women with known cervicovaginal HPV infection status. It is anticipated that the enhanced ELISA conditions will have wide application to a large number of clinical diagnostic assays.

Animals↗

Use of resequencing oligonucleotide microarrays for identification of Streptococcus pyogenes and associated antibiotic resistance determinants.

Group A streptococci (GAS) are responsible for a wide variety of human infections associated with considerable morbidity and mortality. Ever since the first systematic effort by Lancefield to group Streptococcus species by M protein variants, the detection and characterization of Streptococcus by different methods have been an evolving process. The ideal assay for GAS identification not only would provide quick and accurate diagnostic results but also would reveal antibiotic resistance patterns and genotype information, aiding not only in treatment but in epidemiologic assessment as well. The oligonucleotide microarray is a promising new technology which could potentially address this need. In this study, we evaluated the usefulness of oligonucleotide resequencing microarrays for identifying GAS and its associated antibiotic resistance markers. We demonstrated an assay platform that combines the use of resequencing DNA microarrays with either random nucleic acid amplification or multiplex PCR for GAS detection. When detecting Streptococcus pyogenes from coded clinical samples, this approach demonstrated an excellent concordance with a more established culture method. To this end, we showed the potential of resequencing microarrays for efficient and accurate detection of GAS and its associated antibiotic resistance markers with the benefit of sequencing information from microarray analysis.

Anti-Bacterial Agents↗