Elevation of viral load by PCR and use of plasma preparation tubes for quantification of human immunodeficiency virus type 1.

HIV-1 viral load testing from 51 patients was compared in plasma samples simultaneously processed and stored in primary Vacutainer plasma preparation tubes (PPTs) and PPT sample aliquots transferred in secondary tubes before freezing, using RT-PCR quantification with an ultrasensitive method (the Roche AMPLICOR HIV-1 MONITOR). We concluded that freezing the primary sample in the PPT collection tube can artificially elevate the HIV-1 viral load. We therefore store samples as frozen aliquots in a second tube.

Single (B or C), dual (BC or BD) and triple (BCD) viral hepatitis in HIV-infected patients in Madrid, Spain.

INTRODUCTION: There are very limited data about the prevalence of multiple hepatitis virus infections in HIV infected individuals. In HIV uninfected individuals with triple BCD hepatitis, hepatitis D virus (HDV) appears to be the dominant virus. However, in HIV infected patients with triple hepatitis it is not known if HDV replication inhibits hepatitis B virus (HBV) and/or hepatitis C virus (HCV) replication. METHODS: We calculated the prevalence of single (B or C), dual (BC) and triple (BCD) hepatitis in 423 HIV-infected patients with positive HCV serum antibodies and/or positive serum HBsAg. In patients with multiple infections we performed an evaluation of serum markers of HBV, HCV and HDV replication. RESULTS: The prevalence of multiple hepatitis was 4.7% (95% confidence interval, 2.7-6.7%). Multiple hepatitis occurred only among patients who acquired HIV through injection drug use. The most common multiple hepatitis was triple BCD. Patients with hepatitis BC and past or chronic hepatitis D were significantly more likely to have cirrhosis and a negative serum HBeAg and HCV PCR than patients with single hepatitis B or hepatitis C. Patients with chronic hepatitis D showed uniform suppression of HBV and HCV replication markers. CONCLUSIONS: In our geographic area approximately 5% of HIV infected patients with hepatitis suffer multiple hepatitis virus infection. In patients with triple hepatitis BCD virus infection, HDV appears to be the dominant virus causing inhibition of both HBV and HCV replication.

Comparison between sequence analysis and a line probe assay for testing genotypic resistance of human immunodeficiency virus type 1 to antiretroviral drugs.

The purpose of this study was to compare a line probe assay (LiPA) with sequence analysis for the detection of mutations conferring resistance to nucleoside and non-nucleoside inhibitors in human immunodeficiency reverse transcriptase and protease inhibitors. The limitations for interpreting LiPA make it unacceptable for routine clinical practice.

Quantitation of human immunodeficiency virus type 1 RNA loads in cervicovaginal secretions in pregnant women and relationship between viral loads in the genital tract and blood.

The purpose of this study was to analyze the quantitation of the human immunodeficiency virus type 1 RNA (HIV-1 RNA) in the genital tract of HIV-1-infected pregnant women and to evaluate a possible correlation with the viral load in blood plasma (Spearman's rank correlation coefficient). A total of 38 each of cervical, vaginal, and blood samples from 38 women were obtained during the third trimester of pregnancy for quantitation of the HIV-1 RNA load. Viral loads were determined by reverse transcription-polymerase chain reaction. The HIV-1 RNA viral load was detectable in 29 of the 38 (76.3%) blood samples, in 6 of the 38 (15.7%) cervical secretion samples, and in 8 of the 38 (21%) vaginal secretion samples. Overall, the correlation between the HIV-1 RNA viral load in the blood plasma and in cervical secretion samples was 0.51 ( P<0.001). However, the correlation disappeared ( r=0.27) when three patients with high blood plasma viral loads were eliminated from the statistical study. The viral load in the vaginal secretions did not correlate with that in the blood samples ( r=0.26). There were two cases in which HIV-1 RNA was undetectable in the blood and cervix but was detectable in vaginal secretions: one woman had 220 copies/ml and the other 68 copies/ml. These results suggest that pregnant women with undetectable viral loads in blood plasma are still at risk of transmitting the virus vertically during vaginal delivery. Because of this, antiretroviral prophylaxis during vaginal delivery must be administered to HIV-1-infected women and their newborns, regardless of the mother's viral load in plasma. In conclusion, quantification of cervicovaginal levels of HIV-1 may represent a useful tool for assessing the individual risk associated with a vaginal delivery and for guiding decisions about whether a scheduled caesarean should be recommended.

Effects of growth hormone plus a hyperproteic diet on methotrexate-induced injury in rat intestines.

The aim of this study was to determine whether growth hormone treatment reduces injury to the intestinal mucosa induced by methotrexate (MTX). Wistar rats with intestinal injury induced by methotrexate were treated with daily growth hormone, beginning 3 days before MTX treatment until 3 or 4 days after MTX administration. The rats were killed at 3 or 7 days post-MTX administration. The rats were fed with either a normoproteic diet or a hyperproteic diet. Body weight, mortality, bacterial translocation, intestinal morphometry, proliferation and apoptosis and blood somatostatin and IGF-1 were determined. Combined administration of growth hormone and a hyperproteic diet reduces MTX-induced mortality. This effect was accompanied by increased cell proliferation and decreased apoptosis within the crypt. Morphometric data showed complete recovery of the mucosa by day 7 post-MTX administration. These results indicate a synergistic protective action of growth hormone combined with a hyperproteic diet to MTX-induced injury.

[Astrovirus infection in children with gastroenteritis]. / Infección por astrovirus en niños con gastroenteritis.

OBJECTIVE: We report here a 2 year study (1997-1998) of astrovirus infection in children admitted to our hospital. The epidmioligical and clinical features of the infection were also investigated. METHOD: A total of 1.815 stool specimens were tested during a two-year period (january 1997- december 1998). They were collected from children less than 3 years of age admitted to La Paz Hospital (Madrid) with acute gastroenteritis. We used a monoclonal antibody-based enzyme inmunoassay (IDEIA Astrovirus, DAKO) to detect astrovirus antigen in faecal samples. RESULTS: Astroviruses were detected in 79 of the 1.815 analysed samples. This represented an incidence of 4,3%. The main symptom of infection was watery diarrhoea, and it was often associated with vomiting and fever. The majority of astrovirus infection occurred between the coldest months of the year. Children under a year of age were the most susceptible population. There was a considerable number of patients (62%) who develop the infection after 72 hours of admission. CONCLUSION: The rate of detection reported here illustrates the importance of this pathogen in childhood diarrhoea, so we recommend its screening on a routine basis.

[Utility of IgG subclasses in the diagnosis and follow up of hydatidosis]. / Utilidad de las IgG fraccionadas en el diagnóstico y seguimiento de la hidatidosis.

BACKGROUND: The aim of the study is to evaluate the IgG subclasses for the diagnosis and follow-up of hydatidosis disease. MATERIAL AND METHODS: 82 serum of 50 patients previously diagnosed of hydatidosis disease, were studied. This patients were divided into different groups depending on their symptomatology, and 10 serum of healthy individuals were tested for an indirect hemagglutination, and IgG subclasses were studied in all positive cases by an ELISA. RESULTS: IgG1 subclass was positive in 81 out of 82 patients. IgG2 and IgG3 subclasses were positive in 94.4% of symptomatic patients, in 100% of patients with calcified cyst or who had had no radical surgery, and in 0% of patients with calcified cyst or who had had radical surgery. CONCLUSION: IgG1 and IgG4 subclasses can be used together for hydatidosis diagnosis, due to their high specificity and sensibility. IgG4 subclass becomes negative soon if there is a good clinical response by the patient, turns to be positive when there is a disease recrudescence, and keeps positive when residual cyst are left. All this makes IgG4 subclass a good marker in the hydatidosis follow-up.

Etiología de la gastroenteritis aguda en niños hospitalizados menores de 5 años / Etiology of acute gastroenteritis in children under 5 years of age requiring hospitalization

Objetivos: Estudiar la etiología de la gastroenteritis aguda en niños menores de 5 años que precisan hospitalización. Conocer la frecuencia y las características clínicas de la gastroenteritis por rotavirus de origen comunitario y de adquisición nosocomial en nuestro medio. Material y métodos: Se revisaron de forma retrospectiva todos los casos de gastroenteritis aguda en niños menores de 5 años ingresados en el Servicio de Enfermedades Infecciosas durante un periodo de tres años (1995-1997). Las muestras de heces se procesaron en el Servicio de Microbiología para cultivo bacteriológico, búsqueda de parásitos intestinales y detección de antígenos virales. Se incluyeron los niños con gastroenteritis comunitaria ingresados desde el Servicio de Urgencias y los niños con gastroenteritis nosocomial por rotavirus (adquirida después de tres días de estancia hospitalaria o en los diez días posteriores al alta). Resultados: Durante el periodo de estudio, fueron hospitalizados 169 niños menores de 5 años con gastroenteritis aguda, lo que representó el 6,7 por ciento del total de ingresos en nuestra unidad. En 105 pacientes (62 por ciento) se identificó algún enteropatógeno en el coprocultivo. Rotavirus fue el principal patógeno encontrado (44 por ciento), seguido por Salmonella sp. (38 por ciento) y Campylobacter sp. (9,5 por ciento). De forma adicional, 42 niños fueron diagnosticados de gastroenteritis nosocomial por rotavirus, lo que supone que el 47 por ciento de las infecciones por este virus en nuestra unidad fueron de origen nosocomial. La gastroenteritis nosocomial afectó a niños más pequeños (edad media de 6,5 meses frente a 12,7 meses en la adquirida en la comunidad, [p<0,05]), y la clínica fue menos severa, con un porcentaje significativamente menor de vómitos y deshidratación. El tiempo medio de estancia fue superior en los niños con gastroenteritis nosocomial (12 frente a 8 días, [p <0,051). Conclusiones: Rotavirus es la principal causa de hospitalización en niños menores de 5 años con gastroenteritis aguda y, asi mismo, representa el principal enteropatógeno en la gastroenteritis nosocomial en nuestro medio. La gastroenteritis nosocomial por este virus afecta a niños mas pequeños y prolonga de forma significativa la estancia hospitalaria, aunque la clínica es más leve que la de la adquirida en la comunidad (AU)

Incidence of E. coli O157:H7 and other enteropathogens in a Spanish hospital.

From January to December 1997, stools submitted for routine culture were also screened for E. coli O157:H7 to investigate its incidence in our area. Bloody and non-bloody stools were studied. E. coli O157:H7 was not recovered from any of the samples tested. We believe, therefore, that we are in a low rate of infection area, and accordingly do not recommend routine screening of this pathogen.

Evolution of susceptibility of non-typhi Salmonella in a Spanish hospital (1992-1994) and report of a Salmonella ser. Typhimurium isolate resistant to quinolones.

We report the evolution of the antimicrobial resistance of non-typhi Salmonella (1992-1994) in a Spanish hospital in relation with a case of infection due to a fluoroquinolone-resistant strain of Salmonella ser. Typhimurium. None of the isolates were resistant to ciprofloxacin. The resistance to ampicillin has increased from 20.6% (1992) to 29.5% (1994) whereas no significative differences in chloramphenicol and cotrimoxazole susceptibility were noted.

[Rotavirus infection: clinical characteristics and time of elimination of the rotavirus antigen in the feces]. / Infección por rotavirus: características clínicas y tiempo de eliminación del antígeno de rotavirus en heces.

BACKGROUND: A prospective study was performed during three years with 76 children hospitalized with the diagnose of rotaviral gastroenteritis. The aim of our study were to evaluate the clinical features of the rotaviral gastroenteritis, the excretion time of rotavirus in stools and the nosocomial rotavirus infection. METHODS: The detection of the rotaviral antigen in stool was performed using a rapid test based on latex aglutination (Rotalex). For the determination of the excretion time of rotavirus in stools the test was performed daily in 69 children until the results was negative. Furthermore, a clinical study of the patients was carried out. RESULTS: In 39 cases (51.3%) the gastroenteritis was acquired in the community and 37 cases (48.7%) were nosocomially acquired. The most frequent clinical presentation was an acute diarrhea (96%) with vomiting (63.2%) and fever (63.2%). Only three patients did not have diarrhea. The mean duration of diarrhea was between 3-4 days with a highest duration of 10 days. Most of the patients (58%) had an excretion time of rotaviral antigen lower or equal than 3 days, with an extreme value of 14 days. CONCLUSIONS: In our study rotaviral infection occurred mainly in the cooler winter months and in children younger than 6 months of age. A high nosocomial infection level (48.7%) was detected. Vomiting often preceded the onset of diarrhea and fever. The excretion time of rotaviral antigen was independent of the clinical course.

Seroprevalence of toxocariasis in children and a case of VLM.

An epidemiological study on toxocariasis in children was carried out at our hospital (Madrid, Spain) in relation with a case of toxocariasis in a child with chronic hypereosinophilia. The study was based on a positive result of the ELISA test, using excretory-secretory antigen from Toxocara canis. The seroprevalence in children was 1%.

Bacteraemia caused by Campylobacter spp.

The genus Campylobacter has become increasingly recognised as the cause of various infections. Campylobacter jejuni and C coli cause acute gastroenteritis in man all over the world. C jejuni enteritis can lead to bacteraemia, but its actual incidence remains unknown. Seven cases of bacteraemia caused by C jejuni or C coli are reported, from the blood of seven patients: five immune deficient adults; a newborn baby; and a patient who had had abdominal surgery. Patients who develop diarrhoea as a result of Campylobacter infection are at risk of bacteraemia thereafter.

[Evaluation of IgG-IgM antibodies against the Epstein-Barr virus nuclear antigen (EBNA-1) using ELISA]. / Estudio y valoración de anticuerpos IgG-IgM frente al antígeno nuclear del virus de Epstein-Barr (EBNA-1) mediante ELISA.

BACKGROUND: Evaluation of two commercial immunoenzymatic techniques (ELISA) for detection of anti-EBNA-1 IgG and IgM for diagnosis of Epstein-Barr virus mononucleosis. METHODS: Study of anti-EBNA-1 IgG and IgM in infectious mononucleosis and they have been compared with the detection of IgM antibodies to VCA by indirect immunofluorescence as the reference method. RESULTS: The sensitivity of the assay was 87.5%, the specificity was 89.4%, the positive predictive value was 91.4% and the negative predictive value was 85%. CONCLUSIONS: This technique can be useful in laboratories without the necessary equipment and experienced personnel for indirect immunofluorescence. It also can be used as a complementary determination to the anti-VCA IgM in the long lasting infectious mononucleosis, when this marker is negative.