PCR cuantitativa en tiempo real para la amplificación de ADN de Burkholderia mallei: comparación con el método molecular recomendado por la OIE / Quantitative PCR in real time to Burhholderia mallei ADN amplification: A comparison with the molecular method recommended by OIE

Objetivo principal: Comparar dos PCRs en tiempo real cuantitativas para la identificacion de Burkholderia mallei, en terminos de sensibilidad y especificidad analiticas. Metodología: Amplificacion parcial de genes de B. mallei:- orf11 y orf13 del sistema de secreción de tipo III TTS1 del genero Burkholderia mediante qPCR con sondas de hibridacion. - fliP que codifica para la flagelina P de B. mallei mediante qPCR con sonda TaqMan. Calculo de parametros de validez. Resultados: El ensayo desarrollado en el laboratorio obtuvo un limite de deteccion del orden del obtenido con el metodo recomendado por la OIE (70,4 fg/reaccion) y permitio la amplificación especifica de ADN de B. mallei. Conclusión: El metodo desarrollado por el laboratorio de Biologia Molecular del INTA permite una rapida amplificacion de ADN de B.mallei con unas elevadas sensibilidad y especificidad analiticas. Ademas, posibilita la diferenciacion entre B. mallei y B. pseudomallei (AU)

Objective: Comparison of two quantitative real-time PCRs for identification of Burkholderia mallei, on analytical sensitivity and specificity terms. Methods: Partial amplification of Burkholderia mallei gene: - orf11 and orf13 targeting the type III secretion TTS1 system cluster from Burkholderia genus by qPCR using hybridisation probes. - fliP targeting flageling P from B. mallei by qPCR by using TaqMan probe. Validity parameters determination. Results: The duplex test developed by the Molecular Biology Laboratory at INTA obtained a limit of detection similar to that reached by the molecular method recommended by the OIE and permitted the specific amplification of B. mallei DNA. Conclusions: The duplex test developed by the Molecular Biology Laboratory at INTA provides a rapid amplification of B. mallei DNA. It also shows high analytical sensitivity and specificity. Furthermore, this test permits the differentiation between B. mallei and B. pseudomallei (AU)

The transcription factor PLA-1/SKN-1A is expressed in human placenta and regulates the placental lactogen-3 gene expression.

Here we report the selective expression of two POU transcription factor genes, PLA-1 and OCT-1, in human placenta and choriocarcinoma cell lines JAR, JEG-3 and BeWo. Pla-1 protein binds to a POU-consensus DNA sequence in the human placental lactogen-3 (PL-3) promoter and it is capable of trans-activating its transcription up to 18-fold. Other tissue-specific or ubiquitous POU transcription factors such as Pit-1/GHF-1 or Oct-1 showed none or low levels of trans-activation of the PL-3 promoter. In addition, we identified an unique and highly charged region in the N-terminal portion of Pla-1 protein required for full trans-activation of the PL-3 promoter.