Human papillomavirus (HPV) DNA in penile carcinomas in Argentina: Analysis of primary tumors and lymph nodes

Among sexually transmitted diseases, infection by human papillomavirus (HPV) has become one of the most important. On the other hand, though epidemiological data show that some HPV types are closely associated with cervical cancer, few reports have been found with reference to penile carcinoma because of its rare occurrence. The aim of this study was to investigate the relationship between HPV infection and penile cancer in Argentina. A retrospective study was carried out on 38 white men with penile squamous-cell carcinoma. Sixty-five archival fixed biopsies taken from 34 primary penile tumors, 25 nodal metastases, 1 skin “satellite” metastasis and 5 histologically normal lymph nodes were used as specimens. HPV detection and typing were carried out by the polymerase chain reaction (PCR) using generic primers, combined with single-stranded conformational polymorphism (SSCP) analysis. HPV DNA was found in 71% patients, corresponding 81% of them to “high risk” types, with predominance of HPV 18. Both primary tumors and metastases showed concordance of HPV occurrence and type in both lesions. In 3 patients, HPV 16 was detected not only in primary tumors and metastases, but also in histologically normal lymph nodes. Our data indicate that most penile carcinomas in Argentine patients are etiologically related to HPV, especially to “high risk” genital types. The agreement in HPV detection between primary tumors and metastases suggests a potential viral role in tumor progression. HPV detection in otherwise histologically normal lymph nodes might be useful as early marker of a metastatic process.Fil: Picconi, María A.. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C.G. Malbrán”; ArgentinaFil: Eijan, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; ArgentinaFil: Distéfano, Angélica L.. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C.G. Malbrán”; ArgentinaFil: Pueyo, Silvia. Provincia de Buenos Aires. Ministerio de Salud. Hospital Materno Infantil de San Isidro. Servicio de Dermatología; ArgentinaFil: Alonio, Lidia Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C.G. Malbrán”; ArgentinaFil: Gorostidi, Susana. Universidad de Buenos Aires; ArgentinaFil: Teyssié, Angélica R.. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C.G. Malbrán”; ArgentinaFil: Casabone, Bernardo Adalberto. Universidad de Buenos Aires; Argentin

Human Cytomegalovirus: detection of congenital and perinatal infection in Argentina

BACKGROUND: Human cytomegalovirus (CMV) is one of the most commonly found agents of congenital infections. Primary maternal infection is associated with risk of symptomatic congenital diseases, and high morbidity is frequently associated with very low birth weight. Neonates with asymptomatic infection develop various sequelae during infancy. This is the first Argentine study performed in neonates with congenital and postnatal HCMV infection. The purpose of this study was to evaluate the performance of the polymerase chain reaction (PCR) technique with different pairs of primers, to detect cytomegalovirus isolated in tissue cultures and directly in urine and dried blood spot (DBS) specimens. Results were compared with IgM detection. METHODS: The study was performed between 1999 and 2001 on routine samples in the Laboratory. A total of 61 urine and 56 serum samples were selected from 61 newborns/infants, 33 patients whose samples were analyzed during the first two to three weeks of life were considered congenital infections; the remaining 28 patients whose samples were taken later than the third week were grouped as perinatal infections, although only in 4 the perinatal transmission of infection was determined unequivocally Cytomegalovirus diagnosis was made by isolating the virus from urine samples in human foreskin fibroblast cells. Three different primer pairs directed to IE, LA and gB genes were used for the HCMV PCR assay in viral isolates. Subsequently, PCR and nested PCR (nPCR) assays with gB primers were performed directly in urine and in 11 samples of dried blood spot (DBS) on Guthrie Card, these results were then compared with serology. RESULTS: The main clinical manifestations of the 33 patients with congenital infection were purpura, jaundice, hepatomegaly and anaemia. Three patients presented low birth weight as single symptom, 10, intracranial calcifications, and 2, kidney failure. In the 28 patients grouped as with perinatal infection, anaemia, hepatosplenomegaly and enzymatic alteration were predominant, and 4 patients were HIV positive. The primers used to amplify the gB region had a PCR positivity rate of 100%, whereas those that amplified IE and LA regions had a PCR positivity rate of 54% and 61% respectively, in CMV isolates. Amplification by PCR of urine samples (with no previous DNA extraction), using primers for the gB region, detected 34/61 positive samples. Out of the 33 samples from patients with congenital infection, 24 (73%) were positive. When nPCR was used in these samples, all were positive, whereas in the remaining 28 patients, two negative cases were found. Cytomegalovirus DNA detection in 11 samples was also carried out in DBS: 7 DBS samples were positive and 4 were negative. CONCLUSIONS: Primers directed to the gB fragment region were the best choice for the detection of CMV DNA in positive isolates. In congenital infections, direct PCR in urine was positive in a high percentage (73%) of samples; however, in patients grouped as with perinatal infection only 36% of the cases were positive. With n-PCR, total sample positivity reached 97%. PCR technique performed in DBS allowed identifying congenital infection in four patients and to be confirmed in 3. These results show the value of nPCR for the detection of all cases of CMV infection. The assay offers the advantage that it may be performed within the normal working day and provides reliable results in a much shorter time frame than that required for either traditional tissue culture or the shell-viral assay

Molecular variants of human papillomavirus (HPV) types 16 and 18 in adenocarcinomas of the cervix

Fil: Picconi, María Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Alonio, Lidia V. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: García Carrancá, Alejandro. Universidad Nacional Autónoma de México. Departamento de Biología Molecular; México.Fil: Lizano, Marcela. Instituto Nacional de Cancerología; México.Fil: Cervantes Vazquez, Guadalupe. Instituto Nacional de Cancerología; México.Fil: Distéfano, Angélica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Mural, Juan. Hospital Nacional Profesor Alejandro Posadas; Argentina.Fil: Bazan, Juan. Hospital Nacional Profesor Alejandro Posadas; Argentina.Fil: Teyssie, Angelica R. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Los virus Papiloma humano (HPV), en particular los tipos 16 y 18, son considerados carcinógenos humanos, habiéndose demostrado una asociación etiológica entre la infección con estos virus y el desarrollo del cáncer de cuello uterino. El rol viral en el carcinoma escamoso ha sido ampliamente estudiado aunque la información disponible en relación al adenocarcinoma es mucho menor, en parte debido a su baja frecuencia. En este trabajo investigamos la presencia de tipos y variantes intratípicas de HPV en adenocarcinomas de cérvix. Se incluyeron 23 biopsias de archivo, fijadas y embebidas en parafina. La detección y tipificación viral se llevó a cabo mediante PCR genérica y posterior análisis de polimorfismos conformacionales de cadena simple (SSCP). La variabilidad genética se investigó en un fragmento de 450 pb del gen L1, mediante la secuenciación directa post-PCR. Se detectaron 11 muestras positivas para HPV 16 (9 prototipos y 2 variantes: 1 europea y 1 asiática-americana), 10 para HPV 18 (9 prototipos y 1 variante europea), 1 para HPV 31 y 1 negativa. Se confirmó la asociación de HPV de alto riesgo con esta neoplasia, con una alta prevalencia (43%) de HPV 18 pero sin un predominio sobre los demás tipos virales, como fue publicado previamente. La variabilidad demostrada en epítopes de la proteína L1 originaron cambios aminoacídicos que podrían tener implicancias en la repuesta inmune y por lo tanto ser considerados en el diseño de vacunas. Human Papillomavirus (HPV), particularly types 16 and 18, are considered human carcinogens since an etiological association has been demonstrated between these viruses and the development of cervical cancer. While the viral role in squamous carcinoma has been largely studied, the information available on adenocarcinoma is scarce, partly because of its lower frequency. In this paper we investigated the presence of HPV types and intratype variants in adenocarcinomas of the cervix. A total of 23 archive samples, fixed and paraffin embedded biopsies, were included. The detection and viral typing was performed by generic PCR and subsequent single stranded conformational polymorphism analysis (SSCP). Genetic variability was investigated in a 450 bpfragment corresponding to L1 gene by post-PCR direct sequencing. We detected 11 HPV 16 positive samples (9 prototypes and 2 variants: 1 European and 1 Asiatic-American), 10 HPV 18 (9 prototypes and 1 European variant), 1 HPV 31 and 1 negative. The high risk HPV association with this neoplasia was confirmed with a high prevalence (43%) of HPV 18, (but) without predominance over the other types as previously published. The demonstrated variability in L1 protein epitopes originated aminoacidic changes which could have implications on the immune response and therefore should be considered in a vaccine design

Molecular variants of human papillomavirus (HPV) types 16 and 18 in adenocarcinomas of the cervix

Fil: Picconi, María Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Alonio, Lidia V. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: García Carrancá, Alejandro. Universidad Nacional Autónoma de México. Departamento de Biología Molecular; México.Fil: Lizano, Marcela. Instituto Nacional de Cancerología; México.Fil: Cervantes Vazquez, Guadalupe. Instituto Nacional de Cancerología; México.Fil: Distéfano, Angélica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Fil: Mural, Juan. Hospital Nacional Profesor Alejandro Posadas; Argentina.Fil: Bazan, Juan. Hospital Nacional Profesor Alejandro Posadas; Argentina.Fil: Teyssie, Angelica R. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología; Argentina.Los virus Papiloma humano (HPV), en particular los tipos 16 y 18, son considerados carcinógenos humanos, habiéndose demostrado una asociación etiológica entre la infección con estos virus y el desarrollo del cáncer de cuello uterino. El rol viral en el carcinoma escamoso ha sido ampliamente estudiado aunque la información disponible en relación al adenocarcinoma es mucho menor, en parte debido a su baja frecuencia. En este trabajo investigamos la presencia de tipos y variantes intratípicas de HPV en adenocarcinomas de cérvix. Se incluyeron 23 biopsias de archivo, fijadas y embebidas en parafina. La detección y tipificación viral se llevó a cabo mediante PCR genérica y posterior análisis de polimorfismos conformacionales de cadena simple (SSCP). La variabilidad genética se investigó en un fragmento de 450 pb del gen L1, mediante la secuenciación directa post-PCR. Se detectaron 11 muestras positivas para HPV 16 (9 prototipos y 2 variantes: 1 europea y 1 asiática-americana), 10 para HPV 18 (9 prototipos y 1 variante europea), 1 para HPV 31 y 1 negativa. Se confirmó la asociación de HPV de alto riesgo con esta neoplasia, con una alta prevalencia (43%) de HPV 18 pero sin un predominio sobre los demás tipos virales, como fue publicado previamente. La variabilidad demostrada en epítopes de la proteína L1 originaron cambios aminoacídicos que podrían tener implicancias en la repuesta inmune y por lo tanto ser considerados en el diseño de vacunas. Human Papillomavirus (HPV), particularly types 16 and 18, are considered human carcinogens since an etiological association has been demonstrated between these viruses and the development of cervical cancer. While the viral role in squamous carcinoma has been largely studied, the information available on adenocarcinoma is scarce, partly because of its lower frequency. In this paper we investigated the presence of HPV types and intratype variants in adenocarcinomas of the cervix. A total of 23 archive samples, fixed and paraffin embedded biopsies, were included. The detection and viral typing was performed by generic PCR and subsequent single stranded conformational polymorphism analysis (SSCP). Genetic variability was investigated in a 450 bpfragment corresponding to L1 gene by post-PCR direct sequencing. We detected 11 HPV 16 positive samples (9 prototypes and 2 variants: 1 European and 1 Asiatic-American), 10 HPV 18 (9 prototypes and 1 European variant), 1 HPV 31 and 1 negative. The high risk HPV association with this neoplasia was confirmed with a high prevalence (43%) of HPV 18, (but) without predominance over the other types as previously published. The demonstrated variability in L1 protein epitopes originated aminoacidic changes which could have implications on the immune response and therefore should be considered in a vaccine design