Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus

الموضوعات

علوم بحتة وطبيعية

الملخص

Matrix Assisted Laser Desorption Ionization mass spectrometry (MALDI-MS)has been recently introduced in to the field of microbiology. The use of this technique overcomes some of the limitations of current phenotypic and genotypic methods. MALDI-typing of microorganisms is an approach based on the differentiation of MALDI-acquired protein fingerprints. Employed in clinical settings, it allows rapid identification of microorganisms down to the strain level. Leading to high morbidity and mortality rates, antibiotic resistant strains of Staphylococcus aureus have become a worldwide concern. Herein, we carried out a comparative study of 20 variations of an acid/alcohol bacterial protein extraction method using a clinical isolate of S. aureus. Protein fingerprints of these extracts acquired in linear mode (800-20,000 Da) were used for assessment of information content (number of peaks) and identity (size of proteins/peptides, m/z). Two methods were shown to be most efficient for sample preparation, i.e. formic acid/methanol and trifluoroacetic acid/ethanol, yielding 28 peaks each. Proteins obtained by the classic formic acid/ethanol extraction were separated using 2-dimensional electrophoresis (2-DE) yielding more than 50 protein spots. Nine proteins were successfully identified using peptide mass fingerprinting (PMF), two of which are virulence related proteins, Spo VG and endonuclease IV. These proteins are considered prospects for MALDI-typing.

روابط وملفات

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التعريف والنوع

رقم الوثيقة: f2ba2ea7-14f3-49f5-bc0c-de9688459400
رقم العقد: 0
نوع الوسائط: Crawler
نوع المحتوى: الرسائل العلمية
صيغة المصدر: رسائل ماجيستير
نوع الملف: pdf text
أسماء الملفات: 2213210_1.pdf

بيانات النشر

ألقاب المؤلفين: [{"name_ar":" Brigitte Wex","title_ar":"اشراف","title_en":"Supervision"},{"name_ar":"Hussein Hassan El Hage ","title_ar":"اعداد","title_en":"Preparation"}]
اللغة: English

المصدر والدورية

اسم المصدر: Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus

المحتوى والصفحات

عدد الصفحات: 0
كلمات الباحثين: Staphylococcus aureus

إشراف وإعداد

الإشراف: Brigitte Wex
الإعداد: Hussein Hassan El Hage

الاقتباسات الببليوغرافية

APA

Brigitte Wex و Hussein Hassan El Hage . (2013). Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus . أطروحة(رسائل ماجيستير). مدرسة الآداب والعلوم-الجامعة اللبنانية الأميركية. لبنان.

MLA

Brigitte Wex و Hussein Hassan El Hage . Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus . 2013. مدرسة الآداب والعلوم-الجامعة اللبنانية الأميركية، رسائل ماجيستير.