RESUMEN
BACKGROUND: The prevalence of onychomycosis has increased steadily in the past decade. An accurate diagnosis at the outset is important for successful and cost-effective treatment of patients. However, current diagnostic tests for onychomycosis are not rapid, sensitive or specific. OBJECTIVES: To develop a microsatellite-based polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (MS-ELISA) for the detection of Trichophyton rubrum, which is the most common aetiological agent of onychomycosis. METHODS: An archival set of 434 nail and skin specimens from 217 patients was included as the test sample in this study. We also compared MS-ELISA with an earlier published topoisomerase PCR-ELISA (TI-ELISA) using template DNA extracted by another method. RESULTS: The MS-ELISA detected the highest number of positive samples (69%) followed by direct microscopy (56%), TI-ELISA (44%) and fungal culture (30%). When an identical DNA extraction method was applied to 120 specimens, the MS-ELISA proved to be twice as sensitive as the TI-ELISA. CONCLUSIONS: We have optimized a target gene and DNA extraction method for rapid detection of T. rubrum onychomycosis.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Onicomicosis/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Trichophyton/aislamiento & purificación , Anticuerpos Antifúngicos/sangre , ADN de Hongos/análisis , Reacciones Falso Positivas , Humanos , Onicomicosis/microbiología , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Trichophyton/genética , Trichophyton/inmunologíaRESUMEN
BACKGROUND: The prevalence of onychomycosis is rising worldwide. Before starting antifungal treatment, an exact mycological diagnosis should be obtained. The current laboratory diagnosis of dermatomycoses is based on the detection of the causative agent by microscopy and culture. These conventional diagnostic methods for fungal infections often are not the best solution because they are time-consuming, cultures are false-negative and direct examination identifies non-vital structures which cannot be used for speciation. PATIENTS AND METHODS: A total of 218 patients presenting in a surgical practice over 3 months with clinical signs of tinea pedis and/or onychomycosis were involved in the prospective study. All patients had predisposing factors for tinea pedis and tinea unguium, such as vascular insufficiency, diabetes mellitus, and leg ulcers. Nail specimens and skin scrapings were investigated for fungi using Blancophor® preparation, and cultured. In addition to conventional diagnostics, PCR (polymerase chain reaction) for detection of dermatophyte DNA was employed. This PCR-Elisa assay is based on the use of specific primers which target the topoisomerase II gene. This allows the highly specific molecular identification of Trichophyton (T.) rubrum, T. interdigitale, and Epidermophyton floccosum directly in clinical samples. RESULTS: 23.9 % of patients were culture-positive for dermatophytes (either T. rubrum, or T. interdigitale). With PCR, dermatophyte DNA either of T. rubrum or T. interdigitale could be detected in nail samples and skin scrapings from at least 29.9 % of all patients. Epidermophyton floccosum was not found in this study, neither by cultivation nor by PCR. The diagnostic sensitivity of the PCR-Elisa assay was calculated as 79.0% ; the diagnostic specificity as 85.5 %. CONCLUSION: PCR-Elisa evaluation makes possible a rapid, specific and sensitive diagnosis of dermatophytosis of the nails and skin within 24 (maximal 48) hours with identification of the involved species.
Asunto(s)
Arthrodermataceae/clasificación , Arthrodermataceae/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Tipificación Micológica/métodos , Onicomicosis/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Tiña del Pie/diagnóstico , Anciano , Arthrodermataceae/aislamiento & purificación , Diagnóstico Diferencial , Femenino , Humanos , Masculino , Onicomicosis/microbiología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Tiña del Pie/microbiologíaRESUMEN
The receptor tyrosine kinases (RTKs) epidermal growth factor receptor (EGFR), HER2, HER3 and HER4 are involved in the pathogenesis of multiple human malignant neoplasias. However, their role in the carcinogenesis of basal cell carcinomas (BCC) and squamous cell carcinomas (SCC) remains to be elucidated. In order to further define the role of these RTKs, 56 human skin tissue samples of normal skin, BCC and SCC were studied by conventional and differential and quantitative reverse transcriptase-polymerase chain reaction (rtPCR). EGFR and HER3 were predominantly expressed in the BCCs and SCCs, while HER2 was ubiquitously expressed. HER4 was not expressed in any sample. Since in vitro studies have provided compelling evidence that heterodimer formation of these receptors are associated with different signal transduction processes, coexpression patterns might be decisive for the induction and maintenance of a malignant phenotype. These results confirm this concept: isolated HER2 expression and EGFR/HER2 were predominantly found in normal skin, while HER2/HER3 and the triple expression of EGFR/HER2/HER3 were seen more frequently in the BCCs and SCCs compared with normal skin (50% and 40% compared with 26%, respectively). The activation of HER3, in addition to EGFR and HER2, might therefore be associated with the malignant phenotype. However, due to the small numbers in this study, further confirmation of the patterns is needed.
Asunto(s)
Carcinoma Basocelular/diagnóstico , Carcinoma de Células Escamosas/diagnóstico , Receptores ErbB/metabolismo , Genes erbB/fisiología , Neoplasias Cutáneas/diagnóstico , Biopsia/métodos , Ensayo de Inmunoadsorción Enzimática , Regulación Neoplásica de la Expresión Génica/fisiología , Genes erbB-2/fisiología , Humanos , Receptor ErbB-2/metabolismo , Receptor ErbB-4 , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: Nitric oxide (NO) has antithrombotic properties by regulating platelet function, whereas direct effects on plasmatic coagulation are rarely described. In sepsis and inflammation, when synthesis of NO, oxygen radicals and toxic metabolites is crucial, the expression of tissue factor (TF) on monocytes stimulated by lipopolysaccharides (LPS) induces intravascular coagulation. This study was performed to examine the influence of NO and the NO-dependent metabolite peroxynitrite on LPS-induced TF expression and activity in human monocytes. DESIGN: Experimental study. SETTING: Laboratory for cell biology. METHODS: Human peripheral blood mononuclear cells were isolated from buffy coats by gradient centrifugation. The NO-releasing compounds SIN1 and NOC18 were used under different conditions. TF antigen was assayed by flow cytometry, and its activity by a clotting assay. TF-mRNA was measured by reverse transcriptase polymerase chain reaction (RT-PCR-ELISA). MEASUREMENTS AND RESULTS: Whereas NOC18, a pure NO donor, had no effect, SIN1, releasing both NO and superoxide (O2-), reduced TF expression and activity in a dose- and time-dependent manner; superoxide dismutase (SOD) reversed the SIN1-mediated effect. Adding the O2(-)-deliberating system hypoxanthin/xanthin oxidase (which had no significant effect per se) to NOC18, or using the NO and O2- reaction product peroxynitrite resulted in a reduction of TF expression. RT-PCR-ELISA indicated upregulation of TF-mRNA by SIN1 with a peak at 500 microM; higher doses had less effect. CONCLUSION: These data demonstrate an influence of NO on LPS-induced TF expression in monocytes by prior formation of peroxynitrite; furthermore, the balance between NO and O2- seems to play a crucial role.
Asunto(s)
Depuradores de Radicales Libres/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Monocitos/efectos de los fármacos , Nitratos/farmacología , Óxido Nítrico/farmacología , Oxidantes/farmacología , Tromboplastina/efectos de los fármacos , Tromboplastina/genética , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Citometría de Flujo , Depuradores de Radicales Libres/metabolismo , Humanos , Lipopolisacáridos , Monocitos/inmunología , Monocitos/metabolismo , Óxido Nítrico/metabolismo , Tromboplastina/análisis , Tromboplastina/metabolismoRESUMEN
OBJECTIVE: To compare the results obtained by four different techniques for the detection of Chlamydia trachomatis in the male genital tract. DESIGN: Prospective study. SETTING: Andrology unit of a university hospital. PATIENTS: Male infertility patients. INTERVENTIONS: Analysis of semen samples and urethral swabs for the presence of C. trachomatis by recombinant antibody-enzyme-linked immunosorbent assay (rELISA), polymerase chain reaction (PCR), antigen-enzyme immunoassay (EIA) and McCoy cell culture. MAIN OUTCOME MEASURE: Detection of C. trachomatis. RESULTS: In 57 of 205 semen samples (27.8%) immunoglobulin A-antibodies against C. trachomatis were found. In contrast, only 1 of 56 semen samples (1.8%) was positive for C. trachomatis-DNA by PCR, only 1 of 139 semen samples (0.7%) was positive by antigen-EIA, and only 4 of 173 urethral swabs (2.3%) grew C. trachomatis in cell culture. CONCLUSIONS: The discrepancy of positive results found by the antibody-rELISA and direct methods for the detection of C. trachomatis indicates successful eradication of the microorganism in > 90% of antibody-positive men. Therefore, detection of antibodies against C. trachomatis in seminal plasma appears to be of limited diagnostic value.
Asunto(s)
Chlamydia trachomatis/aislamiento & purificación , Técnicas para Inmunoenzimas/clasificación , Reacción en Cadena de la Polimerasa , Semen/microbiología , Uretra/microbiología , Secuencia de Bases , Células Cultivadas , Humanos , Masculino , Sondas Moleculares/genética , Datos de Secuencia Molecular , Estudios Prospectivos , Uretra/patologíaRESUMEN
U. urealyticum with 15.8% and C. trachomatis antibodies with 15.4% were the most prevalent microbiological findings in 209 male infertility patients. The inflammatory marker granulocyte-elastase was significantly increased in men with C. trachomatis; these men also showed significantly decreased citric acid levels indicating inflammatory damage of the prostate induced by C. trachomatis.
Asunto(s)
Infecciones por Chlamydia , Chlamydia trachomatis , Enfermedades de los Genitales Masculinos/microbiología , Infertilidad Masculina/microbiología , Semen/microbiología , Anticuerpos Antibacterianos/análisis , Antígenos Bacterianos/análisis , Chlamydia trachomatis/inmunología , Citratos/metabolismo , Ácido Cítrico , Eyaculación , Escherichia coli/aislamiento & purificación , Humanos , Inflamación , Elastasa de Leucocito , Masculino , Elastasa Pancreática/metabolismo , Semen/fisiología , Ureaplasma/aislamiento & purificaciónRESUMEN
Three hundred eight-nine healthy, infertile patients were studied to determine the effects of inflammation on genital tract organs. Clinically silent inflammation was diagnosed by measuring polymorphonuclear granulocyte (PMN) elastase in semen. Seminal vesicle, prostate, and epididymis functions were assessed by measuring fructose, citric acid, and neutral alpha-glucosidase in semen. There was a significant relationship between high PMN elastase levels and low citric acid levels in semen; fructose and neutral alpha-glucosidase were not related to PMN elastase. Semen samples with increased PMN elastase levels (greater than 250 and greater than 1,000 ng/ml) showed a high incidence of pathologic citric acid levels (67% and 73%, respectively). These biochemical data indicate that the prostate is the main target in clinically silent male genital tract inflammation.
Asunto(s)
Biomarcadores/química , Enfermedades de los Genitales Masculinos/patología , Genitales Masculinos/patología , Semen/química , Citratos/análisis , Ácido Cítrico , Epidídimo/química , Epidídimo/metabolismo , Epidídimo/fisiología , Fructosa/análisis , Genitales Masculinos/metabolismo , Genitales Masculinos/fisiología , Humanos , Inflamación/patología , Masculino , Neutrófilos/enzimología , Elastasa Pancreática/análisis , Próstata/química , Próstata/metabolismo , Próstata/fisiología , Vesículas Seminales/química , Vesículas Seminales/metabolismo , Vesículas Seminales/fisiología , alfa-Glucosidasas/análisisRESUMEN
Seven Georgian male soldiers (19-25 years old) had accidentally been exposed to radiation by Cs-137 between April 1996 and May 1997. No information about the exact time and duration of exposure was available. All patients presented with the subacute stage of Cutaneous Radiation Syndrome with deep painful ulcers on different body sites, predominantly on the legs. Semen analyses showed complete azoospermia in 4 patients, with elevated follicle-stimulating hormone (FSH) in 3 and elevated luteinizing hormone (LH) in 2 of them. One patient had severe oligozoospermia of 7 million sperm per mL, with normal sperm motility and morphology; his FSH and LH levels were elevated. One patient had complete normozoospermia, and the seventh patient had polyzoospermia of 340 million per mL; both of these patients had normal serum hormone levels. Only the patient with oligozoospermia reported a history of delayed testicular descent; his physical examination showed relatively soft and small testicles and a varicocele with considerable reflux. The physical andrological examinations were normal in the other 6 patients. It is very likely that the azoospermia in the 4 patients can be attributed to the radiation accident. In conclusion, it is essential to perform andrological examinations in patients who have been exposed to radiation even if there are only cutaneous injuries detectable, as a high percentage of them can show azoospermia.
Asunto(s)
Radioisótopos de Cesio/efectos adversos , Oligospermia/etiología , Liberación de Radiactividad Peligrosa , Radiodermatitis/etiología , Adulto , Infecciones Bacterianas/diagnóstico , Hormona Folículo Estimulante/sangre , Georgia (República) , Humanos , Hormona Luteinizante/sangre , Masculino , Medicina Militar , Examen Físico , Radiodermatitis/diagnóstico por imagen , Radiodermatitis/cirugía , Úlcera Cutánea/diagnóstico por imagen , Úlcera Cutánea/etiología , Úlcera Cutánea/cirugía , Colgajos Quirúrgicos , Ultrasonografía DopplerRESUMEN
BACKGROUND: PCR-based detection of microorganisms is widely used for diagnostic purposes. Most routine PCR applications do not control for inhibition of PCR, thus leading to false-negative results. METHODS AND RESULTS: One hundred eighteen swab samples obtained from skin and mucosa were investigated for the presence of herpes simplex virus (HSV), varicella-zoster virus (VZV), and the control gene betaglobin by internally controlled PCR with purified and unpurified DNA in parallel. With unpurified DNA, inhibition of PCR was detected in 23% of betaglobin PCRs, 25% of VZV PCRs, and 16% of HSV PCRs versus 3% each for purified DNA. Approximately 20% of the samples with positive results for HSV or VZV had negative or inhibited results using unpurified DNA. CONCLUSION: These results indicate that PCR from clinical swab specimens should be performed exclusively with internal controls because the positive control alone cannot exclude PCR inhibition in individual samples. Purification of DNA will decrease, but not exclude, PCR inhibition.
Asunto(s)
Herpesvirus Humano 3/genética , Herpesvirus Humano 3/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Simplexvirus/genética , Simplexvirus/aislamiento & purificación , Conjuntiva/virología , ADN Viral/aislamiento & purificación , Reacciones Falso Negativas , Herpes Simple/diagnóstico , Herpes Zóster/diagnóstico , Humanos , Mucosa Bucal/virología , Control de Calidad , Piel/virología , Uretra/virologíaRESUMEN
Detection of localized, clinically atypical cutaneous infections with varicella zoster virus (VZV) has proven difficult, as serum antibody tests sometimes are not sensitive and specific enough for that purpose. Therefore immunofluorescence and an internally controlled PCR for VZV are compared for sensitivity. Detection of PCR products was done by ELISA, and if positive, additionally by agarose gel electrophoresis. Of 60 samples 44 were PCR-positive by ELISA (44 = 100%), of which 37 (84%) were also positive on the agarose gel. Thirty-four samples (77%) were positive by immunofluorescence. No sample was positive by immunofluorescence and negative by PCR. A combination of immunofluorescence and PCR with agarose gel analysis detected 42 samples out of 44 positive by PCR ELISA (95%). These results demonstrate that immunofluorescence is a suitable, fast and inexpensive method for routine diagnostics. Additional sensitivity can be achieved by screening immunofluorescence-negative samples by PCR, which is extremely sensitive but time-consuming and labor-intensive.
Asunto(s)
Herpes Zóster/diagnóstico , Herpesvirus Humano 3/aislamiento & purificación , Enfermedades Cutáneas Virales/diagnóstico , Antígenos Virales/aislamiento & purificación , Cartilla de ADN , Electroforesis en Gel de Agar , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente/normas , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/inmunología , Humanos , Reacción en Cadena de la Polimerasa/normas , Valor Predictivo de las Pruebas , Sensibilidad y EspecificidadAsunto(s)
ADN Viral/análisis , Herpesvirus Humano 8/genética , Pénfigo/genética , Piel/química , Alemania , HumanosAsunto(s)
Infecciones por Herpesviridae/epidemiología , Herpesviridae/aislamiento & purificación , Infertilidad Masculina/virología , Semen/virología , Adulto , Cartilla de ADN , Ensayo de Inmunoadsorción Enzimática , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Distribución Aleatoria , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Shingles are caused by an endogenous or exogenous reinfection with varicella zoster virus (VZV). Up to 50% of individuals with Hodgkin's disease develop herpes zoster; however, no association could be shown between the occurrence of herpes zoster and underlying subclinical malignancies. OBJECTIVE: This study was conducted to investigate whether VZV DNA could be detected by polymerase chain reaction (PCR) in the blood of herpes zoster patients and whether there was an association between VZV viraemia and previous or concurrent neoplasias. METHODS: At least five blood samples from 28 patients with herpes zoster were investigated by internally controlled PCR enzyme-linked immunosorbent assay prior to and during therapy with aciclovir. RESULTS: None of 13 patients, two with a history of neoplasia and two with a neoplasia at the time of the study, showed any signs of viraemia with VZV, and 14 patients had inconsistent viraemia, one with a history of neoplasia and two with neoplasia at the time of the study. In one patient VZV DNA was detected in the blood for 6 days. This patient died soon after from metastatic malignant melanoma. CONCLUSIONS: VZV viraemia may occur during herpes zoster episodes, even in patients without evidence of immunosuppression; however, this viraemia is, in most cases, inconsistent and does not provide any specific information concerning underlying unrecognized malignancies.
Asunto(s)
ADN Viral/análisis , Herpes Zóster/diagnóstico , Herpesvirus Humano 3/aislamiento & purificación , Huésped Inmunocomprometido , Neoplasias/diagnóstico , Neoplasias/inmunología , Reacción en Cadena de la Polimerasa/métodos , Viremia/diagnóstico , Aciclovir/administración & dosificación , Adulto , Anciano , Secuencia de Bases , Femenino , Herpes Zóster/sangre , Herpes Zóster/tratamiento farmacológico , Herpes Zóster/epidemiología , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Neoplasias/epidemiología , Pronóstico , Estudios Prospectivos , Factores de Riesgo , Muestreo , Sensibilidad y Especificidad , Viremia/epidemiologíaRESUMEN
Occupational contact dermatitis in hair dressers and beauticians has increased in importance in the past years. Type IV-allergies against glyceryl monothioglycate components of permanent waves are most common. Other occupational allergens include bleach components such as ammonium persulfate and hair dye ingredients such as p-phenylenediamine (PPD) and p-toluylene-diamine (PTD) base. Allergies to hair dyes in customers of hair dressers have rarely been observed. Two female patients developed allergic contact dermatitis of the scalp and face after repeated use of Polycolor intensivtönung schwarz and of Movida color. We also review the current literature on type IV-allergies to components of hair dressing products components.
Asunto(s)
Industria de la Belleza , Dermatitis Alérgica por Contacto/etiología , Dermatosis Facial/etiología , Tinturas para el Cabello/efectos adversos , Adulto , Angioedema/diagnóstico , Dermatitis Alérgica por Contacto/diagnóstico , Diagnóstico Diferencial , Femenino , Humanos , Pruebas del ParcheRESUMEN
The purpose of the study was to identify semen parameters relevant for in vitro fertilization (IVF). A total of 151 ejaculates from 130 patients were analyzed. There was a significant association between successful IVF, high sperm concentration and good sperm motility. Progressive sperm motility correlated best with successful fertilization of oocytes (r = +0.427) and was the only parameter significantly linked to pregnancy. Very low values were sufficient for fertilization in vitro: a sperm concentration of 4.5 x 10(6)/ml and a progressive sperm motility of 20%. The results of analysis of fructose, citrate and neutral alpha-glucosidase as markers of seminal vesicle, prostate and epididymal functions were not different in fertilizing and non-fertilizing semen samples. Moderate increases in granulocyte elastase levels, between 250 and 1000 ng/ml, did not influence IVF. However, two patients with genital tract inflammation (PMN-elastase > 1000 ng/ml) did not fertilize despite excellent semen parameters. These results stress the importance of andrological factors for in vitro fertilization outcome.
Asunto(s)
Fertilización In Vitro , Recuento de Espermatozoides , Motilidad Espermática/fisiología , Interacciones Espermatozoide-Óvulo/fisiología , Donantes de Tejidos , Adulto , Citratos/metabolismo , Ácido Cítrico , Femenino , Fructosa/metabolismo , Granulocitos/enzimología , Humanos , Masculino , Persona de Mediana Edad , Elastasa Pancreática/metabolismo , Embarazo , Resultado del Tratamiento , alfa-Glucosidasas/metabolismoRESUMEN
BACKGROUND: Kaposi's sarcoma occurs in patients seropositive and seronegative for the human immunodeficiency virus (HIV) and has been associated with human herpes virus 8 (HHV8). The purpose of this study was to determine and to compare the amount of HHV8 DNA in formalin-fixed tissue sections of Kaposi's sarcoma. METHODS: From 27 biopsies of Kaposi's sarcoma patients, tissue sections were taken and deparaffinized. Four patients were HIV seronegative and 13 were HIV seropositive. After extraction of DNA copy numbers of HHV8 and beta-globin were determined in every sample by quantitative PCR ELISA using an internal quantitation standard. Results were expressed as HHV8 per beta-globin. RESULTS: No significant differences were found between biopsies from HIV-positive and HIV-negative patients (14.8+/-19.6 HHV8 per 1000 beta-globin in HIV-positive versus 18.0+/-23.5 in HIV-negative patients). CONCLUSIONS: These data suggest that HHV8 viral load in Kaposi's sarcoma is relatively low and does not differ in HIV-positive and HIV-negative samples. The importance of viral load determination for prognosis or treatment monitoring remains to be elucidated.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/patología , Infecciones Oportunistas Relacionadas con el SIDA/virología , Herpesvirus Humano 8/genética , Sarcoma de Kaposi/patología , Sarcoma de Kaposi/virología , Biopsia , ADN Viral/análisis , Ensayo de Inmunoadsorción Enzimática , Herpesvirus Humano 8/aislamiento & purificación , Humanos , Adhesión en Parafina , Reacción en Cadena de la Polimerasa , Carga ViralRESUMEN
BACKGROUND: Monitoring viral load in blood has already been introduced into clinical routine for human immunodeficiency virus and hepatitis C virus. OBJECTIVE: This study was conducted to monitor the decline of herpes simplex (HSV) viral load in the blood of a patient with gingivostomatitis herpetica prior and during acyclovir therapy. METHODS: Analysis was done by quantitative PCR ELISA using an internal quantitation standard. RESULTS: Copy numbers were 66/microl blood prior to therapy, 60 during oral medication with valaciclovir, 97 and 72 copies/microl blood during the first 2 days of intravenous acyclovir therapy, followed by a sharp decline to 8 and 9 copies on days 3 and 4. During the following days, HSV was no longer detectable. CONCLUSION: As this quantitative approach can be easily adjusted to any other PCR, it provides a reliable, easy-to-apply method for monitoring therapy, also during new antiviral clinical trials.
Asunto(s)
Aciclovir/uso terapéutico , Antivirales/uso terapéutico , ADN Viral/sangre , Herpes Simple/tratamiento farmacológico , Simplexvirus/efectos de los fármacos , Adulto , Ensayo de Inmunoadsorción Enzimática , Herpes Simple/sangre , Herpes Simple/virología , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Simplexvirus/genética , Carga ViralRESUMEN
BACKGROUND: Detection of cutaneous infections with herpes simplex virus (HSV) has proven difficult, as serum antibody tests sometimes are not sensitive and specific enough for that purpose. OBJECTIVE: This study was conducted to compare the sensitivity for detection of HSV of an immunofluorescence method (Syva Microtrak) and an internally controlled PCR. METHODS: Cutaneous swabs from skin lesions were analysed by immunofluorescence separately for HSV types 1 and 2 and by competitive PCR. Detection of PCR products was done by ELISA, if positive additionally by agarose gel electrophoresis. RESULTS: Of 79 samples 34 were PCR-positive by ELISA (34 = 100%), of which 23 (68%) were also positive on the agarose gel. Eleven samples (32%) were positive by immunofluorescence. No sample was positive by immunofluorescence and negative by PCR. CONCLUSIONS: These results demonstrate that immunofluorescence using Syva Microtrak is not suitable for exclusion of herpes simplex virus infection as sensitivity was only 32%. However, as immunofluorescence is cheaper and faster than PCR, first screening can be done with immunofluorescence, and negative samples can be investigated by PCR to finally prove or exclude the presence of HSV DNA.
Asunto(s)
Técnica del Anticuerpo Fluorescente , Herpes Simple/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Simplexvirus/aislamiento & purificación , Secuencia de Bases , Técnicas de Cultivo , ADN Viral/análisis , Electroforesis en Gel de Agar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Tripe palms is a descriptive term for a cutaneous paraneoplastic keratoderma. Tripe palms are frequently associated with gastric and pulmonary carcinoma. The pathogenetic mechanism remains unknown. OBJECTIVE: To determine the influence of receptor tyrosine kinases, which are both expressed in pulmonary carcinomas and in human skin, we performed expression studies on epidermal growth factor receptor (EGFR), HER2, HER3 in a skin sample of tripe palms obtained from a patient with non-small-cell lung cancer with lymph node involvement. Two months after diagnosis, the patient had developed palmoplantar 'tripe palms'. Additionally, the expression of SRC, c-myc and p16/ CDKN2 were studied. METHOD: Conventional reverse-transcription polymerase chain reaction was performed on a tissue sample of tripe palms. RESULTS: Weak expression of HER2 and of p16/CDKN2 was found. EGFR, HER3, c-myc and SRC were not expressed. CONCLUSION: Receptor tyrosine kinases of subclass I, the tyrosine kinase SRC and the oncogene c-myc play no major role in the pathogenesis of this case of tripe palms.