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The role of the pediatric nurse in providing laboratory newborn screening
STEINBAUEROVÁ, Julie
This bachelor thesis focuses on the role of the pediatric nurse in newborn laboratory screening. The pediatric nurse has an indispensable role in the entire process, educating parents prior to collection, obtaining informed consent from parents prior to testing, preparing supplies, performing the actual collection, checking and preparing the collected specimens for shipment to the screening laboratory. Although the thesis is primarily focused on maternal education prior to collection and possible errors during collection that may negatively affect the results of the examination, the theoretical and practical part of the thesis also covers other topics related to newborn laboratory screening, not least the description of the methodology of the correct collection procedure and subsequent verification of knowledge of its principles in a research investigation with nurses. The first objective was to describe the process of maternal education by the pediatric nurse prior to collection. The second objective was to determine the knowledge of pediatric nurses about the negative impact of substandard blood collection on screening outcome. The research part of the study was based on qualitative research, using the interview method. Data collection was carried out using the technique of in-depth semi-structured interviews with nurses from neonatology departments of selected Prague hospitals. Data analysis was done by pencil paper method, then the results were divided into three categories and described in diagrams. The survey found that nurses perceive the language barrier as the biggest difficulty in educating mothers before delivery, but they have resources available to help them overcome this barrier in practice. It was also found that nurses are well informed and have sufficient knowledge about the principles of dry drop blood collection. The most common errors in collection were described by nurses as insufficient or incorrectly filled in targets on the filter paper and illegible data on the screening card. This work can be used to educate pediatric nurses, and recommendations for practice include focusing on non-pharmacological methods of procedural pain relief for newborns during collection and checking specimens before sending them to screening laboratories to avoid unnecessary repeat collections.
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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Utilization of LC-MS/MS in diagnosis of congenital adrenal hyperplasia
Grúlová, Kristýna ; Kozlík, Petr (advisor) ; Jelínek, Ivan (referee)
Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease that causes a disorder of steroidogenesis in the adrenal cortex. This disease is a part of a panel of diseases searched in preclinical nationwide neonatal screening. The methodology is based on measuring the concentration of 17-hydroxyprogesterone (17-OHP) in a dried blood spot using fluorescence immunoassay (FIA). However, this determination is not entirely specific and generates a high rate of false positive results (up to 4.3 %). In this diploma thesis the LC-MS / MS method was developed. This method measures selected steroid hormones involved in cortisol metabolism with respect to the diagnosis of CAH disease. The method was validated and applied to clinical samples, it identified CAH patients from negative controls and significantly reduced the false positivity of neonatal screening results. Compared to the FIA results, the LC-MS / MS method reduced false positivity up to 50 % by evaluating the concentration of 17-OHP. Moreover, by extending the diagnostic algorithm with other measured markers, the reduction was enhanced up to 98%. The developed method is also applicable for the measurement of serum and plasma samples, respectively, and has become a part of the confirmation tests for suspected CAH screening findings. Key...
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef ; Jelínek, Ivan (advisor) ; Čabala, Radomír (referee) ; Cvačka, Josef (referee)
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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