Experimental transmission of enzootic nasal adenocarcinoma in sheep.

Enzootic nasal adenocarcinoma (ENA) is a contagious neoplasm of the secretory epithelial cells of the nasal mucosa of sheep and goats. It is associated with the betaretrovirus, enzootic nasal tumor virus (ENTV), but a causative relationship has yet to be demonstrated. In this study, 14-day-old lambs were experimentally infected via nebulization with cell-free tumor filtrates derived from naturally occurring cases of ENA. At 12 weeks post-infection (wpi), one of the five infected lambs developed clinical signs, including continuous nasal discharge and open mouth breathing, and was euthanized. Necropsy revealed the presence of a large bilateral tumor occupying the nasal cavity. At 45 wpi, when the study was terminated, none of the remaining infected sheep showed evidence of tumors either by computed tomography or post-mortem examination. ENTV-1 proviral DNA was detected in the nose, lung, spleen, liver and kidney of the animal with experimentally induced ENA, however there was no evidence of viral protein expression in tissues other than the nose. Density gradient analysis of virus particles purified from the experimentally induced nasal tumor revealed a peak reverse transcriptase (RT) activity at a buoyant density of 1.22 g/mL which was higher than the 1.18 g/mL density of peak RT activity of virus purified from naturally induced ENA. While the 1.22 g/mL fraction contained primarily immature unprocessed virus particles, mature virus particles with a similar morphology to naturally occurring ENA could be identified by electron microscopy. Full-length sequence analysis of the ENTV-1 genome from the experimentally induced tumor revealed very few nucleotide changes relative to the original inoculum with only one conservative amino acid change. Taken together, these results demonstrate that ENTV-1 is associated with transmissible ENA in sheep and that under experimental conditions, lethal tumors are capable of developing in as little as 12 wpi demonstrating the acutely oncogenic nature of this ovine betaretrovirus.

Development of a SYBR Green-based real-time quantitative polymerase chain reaction assay to detect enzootic nasal tumor virus in goats.

Enzootic nasal adenocarcinoma is a contagious respiratory disease in goats that is caused by the enzootic nasal tumor virus 2 (ENTV-2). In order to increase the number of available detection methods for ENTV-2, we developed a SYBR Green real-time polymerase chain reaction (SGrPCR) assay that targets the gag gene of ENTV-2. The low limit of detection of the assay was 3.68 × 101 copies/µL, a hundredfold more sensitive than conventional PCR. The melt curve showed a single sharp melt peak at 83°C, which indicated that there was no non-specific amplification or primer dimer formation. The intra-assay and inter-assay coefficients of variation were 1.58% and 1.82%, respectively. There was no cross-reactivity with closely related goat viruses (i.e., orf virus, peste des petits ruminants virus, goatpox virus, foot-and-mouth disease virus) and endogenous retroviruses. In conclusion, the SGrPCR assay is specific for the gag gene of ENTV-2 and provides a rapid and sensitive approach for detecting ENTV-2 in clinical samples.

L'adénocarcinome nasal enzootique est une maladie respiratoire contagieuse chez les chèvres qui est causé par le virus de la tumeur nasale enzootique 2 (ENTV-2). Afin d'augmenter le nombre de méthodes de détection disponibles pour ENTV-2, nous avons développé un test de réaction en chaîne par polymérase en temps réel SYBR Green (SGrPCR) qui cible le gène gag de ENTV-2. La limite basse de détection du test était de 3,68 × 101 copies/µL, cent fois plus sensible que la PCR conventionnelle. La courbe de fusion montrait un seul pic de fusion net à 83 °C, ce qui indiquait qu'il n'y avait pas d'amplification non spécifique ou de formation de dimère d'amorce. Les coefficients de variation intra-essai et inter-essai étaient respectivement de 1,58 % et 1,82 %. Il n'y avait pas de réactivité croisée avec les virus caprins étroitement apparentés (c'est-à-dire le virus orf, le virus de la peste des petits ruminants, le virus de la variole caprine, le virus de la fièvre aphteuse) et les rétrovirus endogènes. En conclusion, le test SGrPCR est spécifique du gène gag de l'ENTV-2 et fournit une approche rapide et sensible pour la détection d'ENTV-2 dans des échantillons cliniques.(Traduit par Docteur Serge Messier).

Human papilloma virus screening by hybrid capture II in Chinese women of Jiangsu Province.

OBJECTIVE: To survey Human Papilloma Virus (HPV) infection in Chinese Women of Jiangsu Province and discuss the relationship between HPV and the biology of cervical cancer. METHODS: Two thousand, one hundred and fifty-three sexually active women (including 66 cases of cervical cancer) were selected for high-risk human papilloma virus DNA test with Hybrid Capture II (HCII). RESULTS: The overall HPV prevalence was 32.6% (701/2153) with higher positive rates in cervical carcinoma and Cervical Interstitial Neoplasia (CIN) [93.9% and 54.6%] respectively. For women aged 40-59 years, the overall high-risk HPV prevalence was higher than those of other age groups. Compared with CIN I, the positivity rate and viral load of HPV DNA in CIN III is much higher (80.2% vs. 29.9%, 11.89 vs. 0.53). Ninety-four per cent (64/66) of patients with Cervical cancer were detected to be HPV positive. There was no significant difference in HPV DNA among each clinical stage and pathologic grade. But the positive rates and the value of HPV DNA were higher in the patients with cervical interstitial incursion. Eighty per cent of patients (20/25) could become negative within six months after operation. CONCLUSIONS: High-risk HPV DNA test is effective in screening for cervical diseases. HCII is an effective method to detect HPV DNA.

Human papilloma virus screening by hybrid capture II in chinese women of Jiangsu Province / Tamizaje del virus del papiloma humano mediante captura híbrida en mujeres chinas de la Provincia de Jiangsu

OBJECTIVE: To survey Human Papilloma Virus (HPV) infection in Chinese Women of Jiangsu Province and discuss the relationship between HPV and the biology of cervical cancer. METHODS: Two thousand, one hundred and fifty-three sexually active women (including 66 cases of cervical cancer) were selected for high-risk human papilloma virus DNA test with Hybrid Capture II (HCII). RESULTS: The overall HPV prevalence was 32.6% (701/2153) with higher positive rates in cervical carcinoma and Cervical Interstitial Neoplasia (CIN) [93.9% and 54.6%] respectively. For women aged 40-59 years, the overall high-risk HPV prevalence was higher than those of other age groups. Compared with CIN I, the positivity rate and viral load of HPV DNA in CIN III is much higher (80.2% vs 29.9%, 11.89 vs 0.53). Ninety-four per cent (64/66) of patients with Cervical cancer were detected to be HPV positive. There was no significant difference in HPV DNA among each clinical stage and pathologic grade. But the positive rates and the value of HPV DNA were higher in the patients with cervical interstitial incursion. Eighty per cent of patients (20/25) could become negative within six months after operation. CONCLUSIONS: High-risk HPV DNA test is effective in screening for cervical diseases. HC II is an effective method to detect HPV DNA.

OBJETIVO: Investigar la infección por el virus del papiloma humano (VPH) en las mujeres chinas de la Provincia de Jiangsu y analizar la relación entre VPH y la biología del cáncer cervical o del cuello uterino. MÉTODOS: Dos mil ciento cincuenta y tres mujeres sexualmente activas (incluyendo 66 casos de cáncer cervical) fueron seleccionadas para una prueba de ADN con el fin de detectar el virus del papiloma humano de alto riesgo mediante Captura Híbrida 2 (HC2). RESULTADOS: La prevalencia general de VPH fue 32.6% (701/2153), hallándose las tasas positivas más altas en el carcinoma cervical y la neoplasia intersticial cervical (NIC) [93.9% y 54.6%]. Para las mujeres de 40-59 años de edad, la prevalencia general de VPH de alto riesgo fue mayor que para los otros grupos etarios. En comparación con el CIN, la tasa de positividad y la carga viral de ADN del VPH en el CIN es mucho mayor (80.2% vs 29.9%, 11.89 vs 0.53). Se detectó que noventa y cuatro por ciento (64/66) de las pacientes con cáncer del cuello uterino eran VPH positivas. No hubo ninguna diferencia significativa en el ADN del VPH ADN entre cada fase clínica y el grado patológico. No obstante, tanto las tasas positivas como el valor de VPH ADN fueron más altos en las pacientes con incursión intersticial cervical. Ochenta por ciento de las pacientes (20/25) podrían volverse negativas en seis meses tras la operación. CONCLUSIONES: La prueba de ADN para la detección del virus del papiloma humano de alto riesgo es un medio efectivo para el tamizaje de las enfermedades cervicales. El HC2 es un método efectivo para detectar el ADN del VPH.