Rajeev Seth1, Peela Sree Ramchandra Murthy2, Sujatha Sistla3, Mahadevan Subramanian4, Kadhiravan Tamilarasu5. 1. Senior Resident, Department of Clinical Microbiology, Rajiv Gandhi Super Speciality Hospital, Tahirpur, Delhi, India. 2. Ph.D. Student, Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India. 3. Professor and Head, Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India. 4. Professor and Head, Department of Pediatrics, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India. 5. Additional Professor, Department of Medicine, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.
Abstract
INTRODUCTION: Acute bacterial meningitis is one of the major causes of morbidity and mortality in children and geriatric population, especially in developing countries. Methods of identification are standard culture and other phenotypic tests in many resource poor settings. AIM: To use molecular methods for the improvement of aetiological diagnosis of acute pyogenic meningitis in patients. MATERIALS AND METHODS: CSF samples of 125 patients were included for the study. Gram staining and culture were performed according to standard procedures. Antigen was detected using commercial latex agglutination test kit. Multiplex PCR was performed using previously published primers and protocols. Fischer's exact test was used for finding association between presence of the disease and clinical/biochemical parameters, considering two tailed p<0.05 as statistically significant. Sensitivity, specificity, positive and negative predictive values were calculated using Graphpad QuicCalc software. RESULTS: A total of 39 cases (31.2%) were confirmed to be of acute pyogenic meningitis based on biochemical methods. Only 10/39 was positive for the three organisms tested. Multiplex PCR was able to detect one additional isolate each of Streptococcus pneumoniae and Haemophilus influenzae type b. When compared with multiplex PCR as the gold standard, culture and latex agglutination tests had same sensitivity (80%), specificity (100%), PPV (100%) and NPV (97.8%), whereas Gram stain had poor sensitivity (40%) and good specificity (95.6%). Detection rates were higher in multiplex PCR for the two organisms Streptococcus pneumoniae and Haemophilus influenzae type b. CONCLUSION: Multiplex PCR was more sensitive than culture or antigen detection, and employing this assay can significantly increase the speed and accuracy of identification of the pathogen.
INTRODUCTION: Acute bacterial meningitis is one of the major causes of morbidity and mortality in children and geriatric population, especially in developing countries. Methods of identification are standard culture and other phenotypic tests in many resource poor settings. AIM: To use molecular methods for the improvement of aetiological diagnosis of acute pyogenic meningitis in patients. MATERIALS AND METHODS: CSF samples of 125 patients were included for the study. Gram staining and culture were performed according to standard procedures. Antigen was detected using commercial latex agglutination test kit. Multiplex PCR was performed using previously published primers and protocols. Fischer's exact test was used for finding association between presence of the disease and clinical/biochemical parameters, considering two tailed p<0.05 as statistically significant. Sensitivity, specificity, positive and negative predictive values were calculated using Graphpad QuicCalc software. RESULTS: A total of 39 cases (31.2%) were confirmed to be of acute pyogenic meningitis based on biochemical methods. Only 10/39 was positive for the three organisms tested. Multiplex PCR was able to detect one additional isolate each of Streptococcus pneumoniae and Haemophilus influenzae type b. When compared with multiplex PCR as the gold standard, culture and latex agglutination tests had same sensitivity (80%), specificity (100%), PPV (100%) and NPV (97.8%), whereas Gram stain had poor sensitivity (40%) and good specificity (95.6%). Detection rates were higher in multiplex PCR for the two organisms Streptococcus pneumoniae and Haemophilus influenzae type b. CONCLUSION: Multiplex PCR was more sensitive than culture or antigen detection, and employing this assay can significantly increase the speed and accuracy of identification of the pathogen.
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