36 research outputs found
A One-Step Real-Time Multiplex PCR for Screening Y-Chromosomal Microdeletions without Downstream Amplicon Size Analysis
BACKGROUND: Y-chromosomal microdeletions (YCMD) are one of the major genetic causes for non-obstructive azoospermia. Genetic testing for YCMD by multiplex polymerase chain reaction (PCR) is an established method for quick and robust screening of deletions in the AZF regions of the Y-chromosome. Multiplex PCRs have the advantage of including a control gene in every reaction and significantly reducing the number of reactions needed to screen the relevant genomic markers. PRINCIPAL FINDINGS: The widely established "EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions (2004)" were used as a basis for designing a real-time multiplex PCR system, in which the YCMD can simply be identified by their melting points. For this reason, some AZF primers were substituted by primers for regions in their genomic proximity, and the ZFX/ZFY control primer was exchanged by the AMELX/AMELY control primer. Furthermore, we substituted the classical SybrGreen I dye by the novel and high-performing DNA-binding dye EvaGreen™ and put substantial effort in titrating the primer combinations in respect to optimal melting peak separation and peak size. SIGNIFICANCE: With these changes, we were able to develop a platform-independent and robust real-time based multiplex PCR, which makes the need for amplicon identification by electrophoretic sizing expendable. By using an open-source system for real-time PCR analysis, we further demonstrate the applicability of automated melting point and YCMD detection
Association of Spermatogenic Failure with the b2/b3 Partial AZFc Deletion
Infertility affects around 1 in 10 men and in most cases the cause is unknown. The Y chromosome plays an important role in spermatogenesis and specific deletions of this chromosome, the AZF deletions, are associated with spermatogenic failure. Recently partial AZF deletions have been described but their association with spermatogenic failure is unclear. Here we screened a total of 339 men with idiopathic spermatogenic failure, and 256 normozoospermic ancestry-matched men for chromosome microdeletions including AZFa, AZFb, AZFc, and the AZFc partial deletions (gr/gr, b1/b3 and b2/b3)
Towards a unified theory of health-disease: II. Holopathogenesis
Este trabalho apresenta uma abordagem sistemática para a modelagem
de várias classes de enfermidade-moléstia-doença, designada como Holopatogênese. Holopatogênese é definido como um processo de sobre determinação de doenças e condições relacionadas, tomadas como um integral, compreendendo facetas selecionadas da saúde enquanto objeto complexo. Em primeiro lugar, o marco conceitual da Holopatogênese é
apresentado como uma série de três interfaces significativas: biomolecular-
imunolĂłgica, fisiopatolĂłgico-clĂnica e epidemiolĂłgico-ecossocial. Em segundo lugar, proposições derivadas da HolopatogĂŞnese sĂŁo introduzidas a fim de permitir o desenho do complexo doença-enfermidade como uma rede hierárquica de redes. Em terceiro lugar, propõe-se uma formalização de correspondĂŞncias intra e inter nĂvel, processos de sobredeterminação, efeitos e laços componentes da HolopatogĂŞnese. Finalmente, o modelo HolopatogĂŞnese Ă© avaliado como uma patologia teĂłrica compreensiva tomada como passo preliminar para uma teoria unificada de saĂşde-doença