Journal of The Academy of Clinical Microbiologists

ORIGINAL ARTICLE
Year
: 2018  |  Volume : 20  |  Issue : 2  |  Page : 77--83

Nested polymerase chain reaction targeting 16S rRNA gene in diagnosis of acute bacterial meningitis


Harshita Lal1, Abhinav Agarwal2, Ashish Kumar Singh1, Rajniti Prasad2, Anil Kumar Gulati1, Pradyot Prakash1 
1 Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India
2 Department of Paediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India

Correspondence Address:
Dr. Pradyot Prakash
Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi - 221 005, Uttar Pradesh
India

CONTEXT: Diagnosing acute bacterial meningitis (ABM) among children presenting to tertiary health care settings is often difficult because of prior administration of antimicrobials. AIMS: The present study was an attempt to diagnose cases of ABM in children with the help of nested polymerase chain reaction (PCR). SETTINGS AND DESIGN: It is a prospective observational study, in which a total of 84 clinically suspected cases and cerbrospinal fluid (CSF) biochemical parameters suggestive of ABM were included in the study. METHODS AND MATERIAL: CSF samples were subjected to Gram staining, bacterial culture and biochemical identification tests as well as panbacterial nested PCR targeting 16S rRNA gene sequence. Subsequently, a nested multiplex PCR for detection of the three fastidious organisms, viz. Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae, was performed. STATISTICAL ANALYSIS USED: The sensitivity, specificity, PPV, NPV, LR+, and LR− of the tests were calculated. RESULTS: The sensitivity of Gram stain, bacterial culture, and nested PCR targeting 16SrRNA were observed to be 16.21%, 9.45%, and 97.29% respectively. Further, the NPV and LR− the PCR were found to be 83.33 and 0.02 respectively. Species specific nested multiplex PCR was able to detect S. pneumoniae (n = 7), N. meningitidis (n = 2) and H. influenzae (n = 1). CONCLUSIONS: The results indicates that nested PCR targeting 16S rRNA gene may be used in diagnosis of ABM. Further, nested multiplex PCR targeting the three important fastidious bacterial pathogens in ABM cases has showed their presence in our region.


How to cite this article:
Lal H, Agarwal A, Singh AK, Prasad R, Gulati AK, Prakash P. Nested polymerase chain reaction targeting 16S rRNA gene in diagnosis of acute bacterial meningitis.J Acad Clin Microbiol 2018;20:77-83


How to cite this URL:
Lal H, Agarwal A, Singh AK, Prasad R, Gulati AK, Prakash P. Nested polymerase chain reaction targeting 16S rRNA gene in diagnosis of acute bacterial meningitis. J Acad Clin Microbiol [serial online] 2018 [cited 2019 Mar 24 ];20:77-83
Available from: http://www.jacmjournal.org/article.asp?issn=0972-1282;year=2018;volume=20;issue=2;spage=77;epage=83;aulast=Lal;type=0