RESUMEN
Our aim was to investigate if interferon plus ribavirin has any effect on serum HCV quasispecies distribution and the relationship between diversity of HCV quasispecies and treatment response. In all, 21 patients were treated with interferon plus ribavirin for 48 weeks. The presence of HCV quasispecies was determined in serum samples at baseline and at the fourth week of treatment by SSCP analysis of the hypervariable region. SSCP pattern was defined as single or multiple band. A single band was found in six patients and multiple bands in nine. No significant difference was found between SSCP pattern in pretreatment samples and response to the therapy. In none of the patients were observed changes in number of SSCP bands between samples taken at baseline and in the fourth week of the therapy. In conclusion, the complexity of HCV quasispecies before the therapy was not related to treatment response; combined therapy did not affect serum HCV quasispecies.
Asunto(s)
Hepacivirus/aislamiento & purificación , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/virología , Interferón-alfa/administración & dosificación , Ribavirina/administración & dosificación , Adulto , Quimioterapia Combinada , Femenino , Genotipo , Hepacivirus/genética , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo Conformacional Retorcido-Simple , ARN Viral/sangre , Resultado del TratamientoRESUMEN
In this study, we have tested a reverse transcription (RT) nested polymerase chain reaction (nPCR) for detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF), serum samples, and conjunctival swabs (CS) from patients with suspected enterovirus infections. A specific 113-bp fragment was amplified using primers designed based on 5 non coding region of the enterovirus genome. The enterovirus RT-nPCR was able to detect 0.001 plaque forming unit (pfu)/ml. Since no PCR product was detected in each of the CSF, CS and serum samples from patients with proven-non-enterovirus viral infections, this method was found to be specific. EV RNA was detected in all 30 culture-confirmed CSF samples and yielded positive results in 5 out of 7 additional cases of culture-negative CSF samples with other evidences of enterovirus infection. Overall, EV RNA was detected in 95 of the patients with clinical diagnosis of viral central nervous system (CNS) disease and confirmed enterovirus infection. Furthermore, we were able to detect EV RNA in 24 (47) out of 51 CSF samples from patients with clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection. The percentage of positive EV RNA detection in paired CSF and serum samples from 11 patients with an enterovirus isolate in CSF was 100 (11 of 11) and 73 (8 of 11), respectively. In addition, EV-specific IgM was detected in 64 (7 of 11) of the sera tested. The method was also tested against 136 samples of CS from patients with clinical diagnosis of acute hemorrhagic conjunctivitis. Ninety nine of them resulted positive (73), while only 27 (20) had been positive for viral culture. In summary, our study shows the importance of enterovirus RT-nPCR for the diagnosis of enterovirus associated disease in different kind of biological samples and different types of diseases.(AU)
Asunto(s)
Humanos , Animales , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Conjuntivitis Hemorrágica Aguda/virología , Enterovirus/aislamiento & purificación , Meningitis Aséptica/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedad Aguda , Anciano de 80 o más Años , Chlorocebus aethiops , Conjuntivitis Hemorrágica Aguda/sangre , Conjuntivitis Hemorrágica Aguda/líquido cefalorraquídeo , Enterovirus/genética , Células HeLa , Meningitis Aséptica/sangre , Meningitis Aséptica/líquido cefalorraquídeo , Estudios Prospectivos , ARN Viral/sangre , ARN Viral/líquido cefalorraquídeo , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad , Células Tumorales Cultivadas , Células VeroRESUMEN
In this study, we have tested a reverse transcription (RT) nested polymerase chain reaction (nPCR) for detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF), serum samples, and conjunctival swabs (CS) from patients with suspected enterovirus infections. A specific 113-bp fragment was amplified using primers designed based on 5' non coding region of the enterovirus genome. The enterovirus RT-nPCR was able to detect 0.001 plaque forming unit (pfu)/ml. Since no PCR product was detected in each of the CSF, CS and serum samples from patients with proven-non-enterovirus viral infections, this method was found to be specific. EV RNA was detected in all 30 culture-confirmed CSF samples and yielded positive results in 5 out of 7 additional cases of culture-negative CSF samples with other evidences of enterovirus infection. Overall, EV RNA was detected in 95 of the patients with clinical diagnosis of viral central nervous system (CNS) disease and confirmed enterovirus infection. Furthermore, we were able to detect EV RNA in 24 (47) out of 51 CSF samples from patients with clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection. The percentage of positive EV RNA detection in paired CSF and serum samples from 11 patients with an enterovirus isolate in CSF was 100 (11 of 11) and 73 (8 of 11), respectively. In addition, EV-specific IgM was detected in 64 (7 of 11) of the sera tested. The method was also tested against 136 samples of CS from patients with clinical diagnosis of acute hemorrhagic conjunctivitis. Ninety nine of them resulted positive (73), while only 27 (20) had been positive for viral culture. In summary, our study shows the importance of enterovirus RT-nPCR for the diagnosis of enterovirus associated disease in different kind of biological samples and different types of diseases.
Asunto(s)
Humanos , Animales , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Conjuntivitis Hemorrágica Aguda/virología , Enterovirus , Meningitis Aséptica/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedad Aguda , Anciano de 80 o más Años , Chlorocebus aethiops , Conjuntivitis Hemorrágica Aguda/sangre , Conjuntivitis Hemorrágica Aguda/líquido cefalorraquídeo , Enterovirus , Células HeLa , Meningitis Aséptica/sangre , Meningitis Aséptica/líquido cefalorraquídeo , Estudios Prospectivos , ARN Viral , Sensibilidad y Especificidad , Células Tumorales Cultivadas , Células VeroRESUMEN
In this study, we have developed a reverse transcription (RT)-nested polymerase chain reaction (n-PCR) for the detection of mumps virus RNA in cerebrospinal fluid (CSF) from patients with neurological infections. A specific 112-bp fragment was amplified by this method with primers from the nucleoprotein of the mumps virus genome. The mumps virus RT-n-PCR was capable of detecting 0.001 PFU/ml and 0.005 50% tissue culture infective dose/ml. This method was found to be specific, since no PCR product was detected in each of the CSF samples from patients with proven non-mumps virus-related meningitis or encephalitis. Mumps virus RNA was detected in all 18 CSF samples confirmed by culture to be infected with mumps virus. Positive PCR results were obtained for the CSF of 26 of 28 patients that were positive for signs of mumps virus infection (i.e., cultivable virus from urine or oropharyngeal samples or positivity for anti-mumps virus immunoglobulin M) but without cultivable virus in their CSF. Overall, mumps virus RNA was detected in CSF of 96% of the patients with a clinical diagnosis of viral central nervous system (CNS) disease and confirmed mumps virus infection, while mumps virus was isolated in CSF of only 39% of the patients. Furthermore, in a retrospective study, we were able to detect mumps virus RNA in 25 of 55 (46%) CSF samples from patients with a clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection including mumps virus infection. The 25 patients represent 12% of the 236 patients who had a clinical diagnosis of viral CNS infections and whose CSF was examined at our laboratory for a 2-year period. The findings confirm the importance of mumps virus as a causative agent of CNS infections in countries with low vaccine coverage rates. In summary, our study demonstrates the usefulness of the mumps virus RT-n-PCR for the diagnosis of mumps virus CNS disease and suggests that this assay may soon become the "gold standard" test for the diagnosis of mumps virus CNS infection.
Asunto(s)
Enfermedades del Sistema Nervioso Central/virología , Virus de la Parotiditis/aislamiento & purificación , Paperas/líquido cefalorraquídeo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Enfermedades del Sistema Nervioso Central/complicaciones , Ataxia Cerebelosa/inducido químicamente , Ataxia Cerebelosa/complicaciones , Encefalitis/complicaciones , Encefalitis/virología , Síndrome de Guillain-Barré/complicaciones , Síndrome de Guillain-Barré/virología , Humanos , Meningitis Aséptica/complicaciones , Meningitis Aséptica/virología , Paperas/complicaciones , ARN Viral/líquido cefalorraquídeo , Sensibilidad y EspecificidadRESUMEN
In this study, we have tested a reverse transcription (RT) nested polymerase chain reaction (nPCR) for detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF), serum samples, and conjunctival swabs (CS) from patients with suspected enterovirus infections. A specific 113-bp fragment was amplified using primers designed based on 5' non coding region of the enterovirus genome. The enterovirus RT-nPCR was able to detect 0.001 plaque forming unit (pfu)/ml. Since no PCR product was detected in each of the CSF, CS and serum samples from patients with proven-non-enterovirus viral infections, this method was found to be specific. EV RNA was detected in all 30 culture-confirmed CSF samples and yielded positive results in 5 out of 7 additional cases of culture-negative CSF samples with other evidences of enterovirus infection. Overall, EV RNA was detected in 95% of the patients with clinical diagnosis of viral central nervous system (CNS) disease and confirmed enterovirus infection. Furthermore, we were able to detect EV RNA in 24 (47%) out of 51 CSF samples from patients with clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection. The percentage of positive EV RNA detection in paired CSF and serum samples from 11 patients with an enterovirus isolate in CSF was 100% (11 of 11) and 73% (8 of 11), respectively. In addition, EV-specific IgM was detected in 64% (7 of 11) of the sera tested. The method was also tested against 136 samples of CS from patients with clinical diagnosis of acute hemorrhagic conjunctivitis. Ninety nine of them resulted positive (73%), while only 27 (20%) had been positive for viral culture. In summary, our study shows the importance of enterovirus RT-nPCR for the diagnosis of enterovirus associated disease in different kind of biological samples and different types of diseases.
Asunto(s)
Conjuntivitis Hemorrágica Aguda/virología , Enterovirus/aislamiento & purificación , Meningitis Aséptica/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Niño , Preescolar , Chlorocebus aethiops , Conjuntivitis Hemorrágica Aguda/sangre , Conjuntivitis Hemorrágica Aguda/líquido cefalorraquídeo , Enterovirus/genética , Células HeLa , Humanos , Lactante , Meningitis Aséptica/sangre , Meningitis Aséptica/líquido cefalorraquídeo , Persona de Mediana Edad , Estudios Prospectivos , ARN Viral/sangre , ARN Viral/líquido cefalorraquídeo , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad , Células Tumorales Cultivadas , Células VeroRESUMEN
In this study, we have tested a reverse transcription (RT) nested polymerase chain reaction (nPCR) for detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF), serum samples, and conjunctival swabs (CS) from patients with suspected enterovirus infections. A specific 113-bp fragment was amplified using primers designed based on 5 non coding region of the enterovirus genome. The enterovirus RT-nPCR was able to detect 0.001 plaque forming unit (pfu)/ml. Since no PCR product was detected in each of the CSF, CS and serum samples from patients with proven-non-enterovirus viral infections, this method was found to be specific. EV RNA was detected in all 30 culture-confirmed CSF samples and yielded positive results in 5 out of 7 additional cases of culture-negative CSF samples with other evidences of enterovirus infection. Overall, EV RNA was detected in 95
of the patients with clinical diagnosis of viral central nervous system (CNS) disease and confirmed enterovirus infection. Furthermore, we were able to detect EV RNA in 24 (47
) out of 51 CSF samples from patients with clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection. The percentage of positive EV RNA detection in paired CSF and serum samples from 11 patients with an enterovirus isolate in CSF was 100
(11 of 11) and 73
(8 of 11), respectively. In addition, EV-specific IgM was detected in 64
(7 of 11) of the sera tested. The method was also tested against 136 samples of CS from patients with clinical diagnosis of acute hemorrhagic conjunctivitis. Ninety nine of them resulted positive (73
), while only 27 (20
) had been positive for viral culture. In summary, our study shows the importance of enterovirus RT-nPCR for the diagnosis of enterovirus associated disease in different kind of biological samples and different types of diseases.
RESUMEN
There are increasing molecular and clinical evidences that the effects of human immunodeficiency virus (HIV) infection can be modified by coinfection with other viruses. The objective was to investigate the viral interaction between HIV and hepatitis C virus (HCV) after HCV superinfection. A 16 year-old pregnant woman was evaluated because of icteric acute hepatitis. Admission laboratory tests showed the following results: ALT 877 IU/L; AST 1822 IU/L; bilirubin 6.79 mg/dl. Diagnosis of acute HCV was based on detection of serum HCV RNA by PCR and anti-HCV seroconversion. ELISA for anti HIV testing was positive and confirmed by western blot. Serum markers for other viruses were negative. The patient was followed during 19 months; serum samples were taken monthly during this period for detection of plasma HIV and HCV RNA. Levels of plasma HIV-RNA were positive in all samples tested before and after the onset of acute hepatitis C. Six months later and a for two month period, and 13 months later for a period of one month HIV viremia was undetectable; then HIV-RNA in plasma was detectable again. In conclusion, HCV superinfection may have temporarily interfered with HIV replication in our patient. The following observations support our hypothesis: it has been demonstrated that HIV-1 replication is suppressed by HCV core protein which has transcriptional regulation properties of several viral and cellular promoters. Clinical implications of this event are not generally known and the interaction between these two viruses in dual infections is worth considering.
Asunto(s)
Regulación hacia Abajo , Infecciones por VIH/complicaciones , VIH/fisiología , Hepacivirus , Hepatitis C/complicaciones , Complicaciones Infecciosas del Embarazo/virología , Sobreinfección/virología , Replicación Viral , Adolescente , Femenino , Infecciones por VIH/virología , Hepatitis C/virología , Humanos , Embarazo , ARN Viral/análisis , Interferencia Viral/fisiologíaRESUMEN
There are increasing molecular and clinical evidences that the effects of human immunodeficiency virus (HIV) infection can be modified by coinfection with other viruses. The objective was to investigate the viral interaction between HIV and hepatitis C virus (HCV) after HCV superinfection. A 16 year-old pregnant woman was evaluated because of icteric acute hepatitis. Admission laboratory tests showed the following results: ALT 877 IU/L; AST 1822 IU/L; bilirubin 6.79 mg/dl. Diagnosis of acute HCV was based on detection of serum HCV RNA by PCR and anti-HCV seroconversion. ELISA for anti HIV testing was positive and confirmed by western blot. Serum markers for other viruses were negative. The patient was followed during 19 months; serum samples were taken monthly during this period for detection of plasma HIV and HCV RNA. Levels of plasma HIV-RNA were positive in all samples tested before and after the onset of acute hepatitis C. Six months later and a for two month period, and 13 months later for a period of one month HIV viremia was undetectable; then HIV-RNA in plasma was detectable again. In conclusion, HCV superinfection may have temporarily interfered with HIV replication in our patient. The following observations support our hypothesis: it has been demonstrated that HIV-1 replication is suppressed by HCV core protein which has transcriptional regulation properties of several viral and cellular promoters. Clinical implications of this event are not generally known and the interaction between these two viruses in dual infections is worth considering.
RESUMEN
Rubella virus antibodies were measured in 85 sera from pregnant women by using a new latex test, and the results were compared with those obtained by using hemagglutination inhibition. The sensitivity of the latex test was 98.4%, specificity was 66.6% and the predictive value of a positive result was 90%. The latex test is a simple test and has much shorter reaction time than that of the hemagglutination inhibition.
Asunto(s)
Anticuerpos Antivirales/análisis , Pruebas de Inhibición de Hemaglutinación , Pruebas de Fijación de Látex , Complicaciones Infecciosas del Embarazo/diagnóstico , Virus de la Rubéola/inmunología , Rubéola (Sarampión Alemán)/diagnóstico , Femenino , Humanos , Valor Predictivo de las Pruebas , EmbarazoRESUMEN
Rubella virus antibodies were measured in 85 sera from pregnant women by using a new latex test, and the results were compared with those obtained by using hemagglutination inhibition. The sensitivity of the latex test was 98.4
, specificity was 66.6
and the predictive value of a positive result was 90
. The latex test is a simple test and has much shorter reaction time than that of the hemagglutination inhibition.