Comparative Study Using MALDI TOF MS Biotyper, Recent Molecular Assays and Conventional Method for Detecting Specified Microbial Contaminants in topical preparations /
Salma Ashraf Mohammed Elqadee
Comparative Study Using MALDI TOF MS Biotyper, Recent Molecular Assays and Conventional Method for Detecting Specified Microbial Contaminants in topical preparations / دراسة للمقارنة بين استخدام تقنية الإمتصاص الضوئي و الفحوصات الجزيئية الحديثة و الطرق التقليدية للكشف عن البكتيريا المُسبّبة لتلوث المستحضرات الصيدلية الجلدية Salma Ashraf Mohammed Elqadee ; Supervised Mohammad Abdalhaleem Ramadan , Omneya Mohammed Helmy , Omneya Mohammed Helmy , Tariq H. Almorsy - Cairo : Salma Ashraf Mohammed Elqadee , 2019 - 77 P. : charts , photographs ; 25cm
Thesis (M.Sc.) - Cairo University - Faculty of Pharmacy - Department of Microbiology and Immunology
Absence of pathogenic bacteria is a must for passing the microbiological quality control inspection of non-sterile pharmaceutical products. The identification of microbial contaminants by conventional methods is time-consuming and labor-intensive. The overall aim of the study was to test non-conventional methods as polymerase chain reaction (PCR) and Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometery Biotyper (MALDI-TOF) for rapid identification of Pseudomonas aeruginosa and Staphylococcus aureus in topical pharmaceutical preparations. Four different topical preparations and three raw materials were artificially inoculated with tested microorganisms, at an initial cell count of 100 CFU/ml, and were overnight pre-enriched in Tryptic soy broth (TSB). MALDI-TOF was tested for identification of target microorganisms using different sample preparation methods; the direct transfer method using single separated colony of target microorganism and two other methods using the incubated broth directly; Formic acid extraction method and newly proposed method , Direct pellet method. PCR targeting oprL (Pseudomonas aeruginosa) and mRNA nuclease gene (Staphylococcus aureus) was also tested using two DNA extraction methods; Mild lysis method was used for detection of single microbial contaminant, Uniplex PCR while QIAmp DNA minikit was used for detection of both microorganisms, duplex and uniplex PCR
MALDI-TOF Non-sterile PCR
Comparative Study Using MALDI TOF MS Biotyper, Recent Molecular Assays and Conventional Method for Detecting Specified Microbial Contaminants in topical preparations / دراسة للمقارنة بين استخدام تقنية الإمتصاص الضوئي و الفحوصات الجزيئية الحديثة و الطرق التقليدية للكشف عن البكتيريا المُسبّبة لتلوث المستحضرات الصيدلية الجلدية Salma Ashraf Mohammed Elqadee ; Supervised Mohammad Abdalhaleem Ramadan , Omneya Mohammed Helmy , Omneya Mohammed Helmy , Tariq H. Almorsy - Cairo : Salma Ashraf Mohammed Elqadee , 2019 - 77 P. : charts , photographs ; 25cm
Thesis (M.Sc.) - Cairo University - Faculty of Pharmacy - Department of Microbiology and Immunology
Absence of pathogenic bacteria is a must for passing the microbiological quality control inspection of non-sterile pharmaceutical products. The identification of microbial contaminants by conventional methods is time-consuming and labor-intensive. The overall aim of the study was to test non-conventional methods as polymerase chain reaction (PCR) and Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometery Biotyper (MALDI-TOF) for rapid identification of Pseudomonas aeruginosa and Staphylococcus aureus in topical pharmaceutical preparations. Four different topical preparations and three raw materials were artificially inoculated with tested microorganisms, at an initial cell count of 100 CFU/ml, and were overnight pre-enriched in Tryptic soy broth (TSB). MALDI-TOF was tested for identification of target microorganisms using different sample preparation methods; the direct transfer method using single separated colony of target microorganism and two other methods using the incubated broth directly; Formic acid extraction method and newly proposed method , Direct pellet method. PCR targeting oprL (Pseudomonas aeruginosa) and mRNA nuclease gene (Staphylococcus aureus) was also tested using two DNA extraction methods; Mild lysis method was used for detection of single microbial contaminant, Uniplex PCR while QIAmp DNA minikit was used for detection of both microorganisms, duplex and uniplex PCR
MALDI-TOF Non-sterile PCR