*1,3Peletiri, I. C., 1Ikeh, E. I., 2Nna, E., 3Ndike, U. P., 4Usman, Y. B., 5Durfa, L. D., 6Okonkwo, C. N., 7Murtala, R., and 2Nnajide, C. R.
1Department of Medical Microbiology, Faculty of Clinical Sciences, College of Health Sciences, University of Jos, Nigeria
2Safety Molecular Pathology Laboratory, The Molecular Pathology Institute, Enugu, Enugu State, Nigeria
3Medical Microbiology & Parasitology Laboratories, National Hospital, Abuja, Nigeria
4Public Health Laboratory, Ministry of Health, Kebbi, Kebbi State, Nigeria
5Microbiology Laboratory, Plateau State Specialist Hospital, Jos, Nigeria
6Medical Laboratory Department, Specialist Hospital, Sokoto, Sokoto State, Nigeria
7Medical Laboratory Services, Ahmad Sani Yariman Specialist Hospital, Gusau, Zamfara State, Nigeria *Correspondence to: kumochris@hotmail.com
Abstract:
Background: Following an increase in the practice of starting antimicrobial therapy prior to clinical sample collection, the ability to confirm pathogenic microorganisms of bacterial meningitis has decreased by approximately 30%. Culture results may be false negative when fastidious or culture-resistant bacteria are involved or when patient samples are obtained after antimicrobial therapy has started. Molecular diagnosis using PCR can be performed directly on clinical samples after metagenomic DNA (mDNA) extraction not requiring live organisms for a positive result. The specific objectives of this study are to perform mDNA extraction directly from cerebrospinal fluids (CSF) using appropriate spin column method, and to determine the quality of the mDNA elute.
Methodology: Cerebrospinal fluid specimens were collected from 210 patients with suspected acute cerebrospinal meningitis (CSM) in the Federal Capital Territory and some States in Northern Nigeria during the 2017 and 2018 outbreak seasons. Metagenomic DNA was extracted from approximately 200µL of CSF specimens using the Qiagen QIAamp(R) DNA Mini kit specific for bacterial agents only. DNA quality check was performed on all DNA elutes using fluorometric, spectrophotometric and agarose gel electrophoresis methods. Continue reading “Quality of metagenomic DNA extracted for molecular identification of microorganisms from CSF samples of patients with suspected cerebrospinal meningitis in northern Nigeria”