Multiplex PCR detection of bacterial and viral meningitis in Cerebrospinal Fluid

الموضوعات

طب

الملخص

Bacterial meningitis is a serious infection of the fluid in the spinal cord and the fluid that surrounds the brain. A multiplex assay was devised to provide simultaneous detection and discrimination of bacterial and Herpes viruses as pathogenic agents of meningitis. A DNA extraction method that was suitable for both bacterial and viral meningitis was devised; the method was designed to allow concentration of viral particles by ammonium sulfate precipitation, for bacterial pathogens, simultaneous treatment of the sample with 2-mercaptoethanol. The prepared samples were tested for amplifiable bacterial or Herpes DNA. Detection was successful when samples were fresh, or when DNA was correctly preserved. Old samples with crude DNA extracts usually produced negative PCR results indicating the unstable nature of crude DNA extract and old samples. The PCR amplification of bacterial DNA was based on previous studies by Barghouthi (2009) where the universal method bacterial multiplex (G7) was utilized directly. A new multiplex (IM) targeting Human Herpes Viruses 1-6 and 8 was designed in this study. IM multiplex was capable of detecting HHV DNA when extracted from water or several dilutions of adulterated CSF (HHV 3/VZV). Epstein-Barr virus was refractory to detection even when two additional primer pairs were introduced, EBV was undetectable from DNA and sera of patients with mononucleosis. Simultaneous detection of bacterial and VZV virus or CMV was possible when IM was combined with G7 in a single multiplex (IMG7). The results were promising since the combined IMG7 multiplex effectively had detected VZV or CMV DNA extracted from CSF. IMG7 was also efficient in detecting Escherichia coli from adulterated CSF. The results were encouraging with the exception of EBV.

روابط وملفات

التعريف والنوع

رقم الوثيقة

0a597bf1-2439-412a-9983-247fa54e2c89

رقم العقد

0

نوع الوسائط

Crawler

نوع المحتوى

الرسائل العلمية

صيغة المصدر

رسائل ماجيستير

نوع الملف

pdf text

أسماء الملفات

1696615_1.pdf

بيانات النشر

ترجمة العنوان

إستخدام متعددة PCR الكشف عن التهاب السحايا البكتيرية والفيروسية في السائل المخي الشوكي.

ألقاب المؤلفين

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اللغة

English

المصدر والدورية

اسم المصدر

Multiplex PCR detection of bacterial and viral meningitis in Cerebrospinal Fluid

المحتوى والصفحات

عدد الصفحات

0

إشراف وإعداد

الإشراف

سمير البرغوثي

الإعداد

ايمان علي محمد مناصره

الاقتباسات الببليوغرافية