Since 1996, the identification of the HFE gene has enabled DNA testing for hereditary haemochroma... more Since 1996, the identification of the HFE gene has enabled DNA testing for hereditary haemochromatosis (HH). The range of DNA testing available includes: (1) diagnostic, (2) predictive (also called presymptomatic testing) and (3) screening. Access to DNA testing has been facilitated by an Australian Medicare rebate, the first available for genetic disorders. Despite the availability of HFE DNA testing in HH, it remains necessary to interpret results in the context of the clinical picture. Traditional markers based on phenotype (transferrin saturation, ferritin and liver biopsy) are still required in some circumstances. We report our experience with HFE DNA testing using a semi-automated approach, which allows multiplexing for the two common mutations (C282Y and H63D). Screening a cohort of b-thalassaemia major and sickle cell anaemia patients of predominantly Mediterranean origin showed that these individuals do not have the common C282Y mutation. This excluded C282Y as a factor in the pathogenesis of iron overload in these haemoglobinopathies. It also showed that the C282Y mutation is of limited value when investigating HH in certain ethnic groups. An Australian family studied illustrated the relative contribution of C282Y and H63D in iron overload. A recently reported third mutation (S65C) in the HFE gene was detected in a low frequency in the populations tested.
New innovations in the diagnostic laboratory achieve their full potential when they can be automa... more New innovations in the diagnostic laboratory achieve their full potential when they can be automated. Increasingly molecular biology (DNA) techniques are being utilised in traditional pathology disciplines, as well as the more recent ones of cytogenetics and molecular genetics. Molecular biology was first exploited for diagnostic purposes when Southern blotting became established. However the time-consuming nature of the methodology, as well as the skills required, made it difficult for Southern blotting to be used routinely in the service laboratory. Subsequently the invention of PCR facilitated the approach to DNA diagnostics. Today PCR in commercial or home-made kits is used for a range of procedures. The steps required to amplify DNA with PCR can also be fully automated. However the analysis of PCR products, which frequently requires slab gel electrophoresis and toxic chemicals or radioisotopes for visualisation, remains difficult to automate. An alternative way for analysing PCR products is now available through capillary electrophoresis. With this technique, automation can be extended to sample loading, electrophoresis and data analysis. The use of toxic chemicals or radioisotopes can be avoided.
Modern manufacturing facilities for bioproducts are highly automated with advanced process monito... more Modern manufacturing facilities for bioproducts are highly automated with advanced process monitoring and data archiving systems. The time dynamics of hundreds of process parameters and outcome variables over a large number of production runs are archived in the data warehouse. This vast amount of data is a vital resource to comprehend the complex characteristics of bioprocesses and enhance production robustness. Cell culture process data from 108 'trains' comprising production as well as inoculum bioreactors from Genentech's manufacturing facility were investigated. Each run constitutes over one-hundred on-line and off-line temporal parameters. A kernel-based approach combined with a maximum marginbased support vector regression algorithm was used to integrate all the process parameters and develop predictive models for a key cell culture performance parameter. The model was also used to identify and rank process parameters according to their relevance in predicting process outcome. Evaluation of cell culture stage-specific models indicates that production performance can be reliably predicted days prior to harvest. Strong associations between several temporal parameters at various manufacturing stages and final process outcome were uncovered. This model-based data mining represents an important step forward in establishing a process data-driven knowledge discovery in bioprocesses. Implementation of this methodology on the manufacturing floor can facilitate a real-time decision making process and thereby improve the robustness of large scale bioprocesses.
Since 1996, the identification of the HFE gene has enabled DNA testing for hereditary haemochroma... more Since 1996, the identification of the HFE gene has enabled DNA testing for hereditary haemochromatosis (HH). The range of DNA testing available includes: (1) diagnostic, (2) predictive (also called presymptomatic testing) and (3) screening. Access to DNA testing has been facilitated by an Australian Medicare rebate, the first available for genetic disorders. Despite the availability of HFE DNA testing in HH, it remains necessary to interpret results in the context of the clinical picture. Traditional markers based on phenotype (transferrin saturation, ferritin and liver biopsy) are still required in some circumstances. We report our experience with HFE DNA testing using a semi-automated approach, which allows multiplexing for the two common mutations (C282Y and H63D). Screening a cohort of b-thalassaemia major and sickle cell anaemia patients of predominantly Mediterranean origin showed that these individuals do not have the common C282Y mutation. This excluded C282Y as a factor in the pathogenesis of iron overload in these haemoglobinopathies. It also showed that the C282Y mutation is of limited value when investigating HH in certain ethnic groups. An Australian family studied illustrated the relative contribution of C282Y and H63D in iron overload. A recently reported third mutation (S65C) in the HFE gene was detected in a low frequency in the populations tested.
New innovations in the diagnostic laboratory achieve their full potential when they can be automa... more New innovations in the diagnostic laboratory achieve their full potential when they can be automated. Increasingly molecular biology (DNA) techniques are being utilised in traditional pathology disciplines, as well as the more recent ones of cytogenetics and molecular genetics. Molecular biology was first exploited for diagnostic purposes when Southern blotting became established. However the time-consuming nature of the methodology, as well as the skills required, made it difficult for Southern blotting to be used routinely in the service laboratory. Subsequently the invention of PCR facilitated the approach to DNA diagnostics. Today PCR in commercial or home-made kits is used for a range of procedures. The steps required to amplify DNA with PCR can also be fully automated. However the analysis of PCR products, which frequently requires slab gel electrophoresis and toxic chemicals or radioisotopes for visualisation, remains difficult to automate. An alternative way for analysing PCR products is now available through capillary electrophoresis. With this technique, automation can be extended to sample loading, electrophoresis and data analysis. The use of toxic chemicals or radioisotopes can be avoided.
Modern manufacturing facilities for bioproducts are highly automated with advanced process monito... more Modern manufacturing facilities for bioproducts are highly automated with advanced process monitoring and data archiving systems. The time dynamics of hundreds of process parameters and outcome variables over a large number of production runs are archived in the data warehouse. This vast amount of data is a vital resource to comprehend the complex characteristics of bioprocesses and enhance production robustness. Cell culture process data from 108 'trains' comprising production as well as inoculum bioreactors from Genentech's manufacturing facility were investigated. Each run constitutes over one-hundred on-line and off-line temporal parameters. A kernel-based approach combined with a maximum marginbased support vector regression algorithm was used to integrate all the process parameters and develop predictive models for a key cell culture performance parameter. The model was also used to identify and rank process parameters according to their relevance in predicting process outcome. Evaluation of cell culture stage-specific models indicates that production performance can be reliably predicted days prior to harvest. Strong associations between several temporal parameters at various manufacturing stages and final process outcome were uncovered. This model-based data mining represents an important step forward in establishing a process data-driven knowledge discovery in bioprocesses. Implementation of this methodology on the manufacturing floor can facilitate a real-time decision making process and thereby improve the robustness of large scale bioprocesses.
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