Impact of Molecular Method for the Diagnosis of Acute Bacterial Meningitis in a Tertiary Health Care Centre in North India (original) (raw)
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Epidemiology and Infection, 2007
To enhance the detection of bacterial meningitis in an East Asian surveillance study, we employed cerebrospinal fluid (CSF) bacterial culture, latex agglutination (LA) and polymerase chain reaction-enzyme immunoassay (PCR-EIA) testing for Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (Sp). The sensitivity and specificity of CSF PCR-EIA testing was compared to LA and culture. A meningitis case was defined by one positive result for any of the three tests. The sensitivity of H. influenzae CSF PCR-EIA, LA, and culture was 100%, 40 % and 57 . 5% respectively; and for Sp CSF PCR-EIA, LA and culture, the sensitivity was 100 %, 58 . 3% and 66 . 7 %, respectively. Hib and Sp specificity was 100% by each method. CSF PCR-EIA was more sensitive than culture or LA for the detection of Hib and Sp meningitis cases increasing their incidence by 74 % and 50 % compared to culture respectively. CSF PCR-EIA should be included for the detection of bacterial meningitis in surveillance studies.
Infectious Disease Reports, 2021
The rapid identification of bacteria causing meningitis is crucial as delays in the treatment increase mortality rate. Though considered as the gold standard for the laboratory diagnosis of bacterial meningitis, culture might give false negative results in a case of patients under antibiotics prior to lumbar puncture. This study aimed to detect Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae by a multiplex polymerase chain reaction (PCR) in culture-negative cerebrospinal fluid samples collected from clinically suspected meningitis cases attending different hospitals in Kathmandu, Nepal from January 2017 to December 2019. S. pneumoniae, N. meningitidis and H. influenzae were detected in 8.59% (33/384) of the specimens by PCR and 7.55% (29/384) of the specimens by culture. Correlation between culture and PCR of the same sample was good (Spearman’s rho correlation coefficient = 0.932). However, the difference in positivity between culture and PCR was statist...
Efficiency of Real-Time PCR in the Diagnosis of Community-Acquired Bacterial Meningitis in Children
Journal of Microbiology and Infectious Diseases
Objectives: Community-acquired bacterial meningitis (CABM) is a life-threatening condition and remains a public health concern despite various efforts to prevent it. This study aimed to detect the bacteria causing CABM in children by Real-Time PCR. Methods: In total, 178 Cerebrospinal fluid (CSF) samples from suspected meningitis cases were collected and subjected to cell count, biochemical, microbiological, and molecular analysis. Bacteria grown on blood and chocolate agar were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). DNA from CSF was extracted and used to detect bacteria by Real-Time PCR using TaqMan Probe. Results: Fifty (28.09%) patients were diagnosed with confirmed meningitis. Of them, 46 (25.84%) were Real-Time PCR, and four (2.25%) were culture and Real-Time PCR positive. Out of 50 bacteria detected, S. pneumoniae (n=35, 19.7%) was the leading causative bacteria and was followed by H. influenzae (seven, 3.93%)...
Journal of pharmaceutical research international, 2021
Background: Meningitis is a rigorous childhood disease with high morbidity and mortality. It is the main cause of under five mortality in India. Mainly three bacteria Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae are responsible. In low economic set up country like India, documented bacterial meningitis mainly depend on gram staining, cerebrospinal fluid (CSF) culture results or latex agglutination test resulting in less number of positive due to the prior antimicrobial intake which affects culture and latex agglutination test results. This study was taken up rapid and accurate molecular method like RT PCR to diagnose bacterial meningitis in culture-negative CSF samples. Materials and Methods: Fifty culture-negative CSF samples from suspected cases of bacterial meningitis were examined by real-time polymerase chain reaction (real-time PCR) for the presence of lytA, bexA, and ctrA genes specific for Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis respectively. Results: Positive real-time PCR results for Streptococcus pneumoniae were detected in 36 (72%) of culture-negative CSF samples while 10% positive results for Haemophilus influenzae type b. Nine (18%) samples were negative by real-time PCR for all tested organisms.
PloS one, 2016
Neisseria meningitidis (Nm), Haemophilus influenzae (Hi), and Streptococcus pneumoniae (Sp) are the lead causes of bacterial meningitis. Detection of these pathogens from clinical specimens using traditional real-time PCR (rt-PCR) requires DNA extraction to remove the PCR inhibitors prior to testing, which is time consuming and labor intensive. In this study, five species-specific (Nm-sodC and -ctrA, Hi-hpd#1 and -hpd#3 and Sp-lytA) and six serogroup-specific rt-PCR tests (A, B, C, W, X, Y) targeting Nm capsular genes were evaluated in the two direct rt-PCR methods using PerfeCTa and 5x Omni that do not require DNA extraction. The sensitivity and specify of the two direct rt-PCR methods were compared to TaqMan traditional rt-PCR, the current standard rt-PCR method for the detection of meningitis pathogens. The LLD for all 11 rt-PCR tests ranged from 6,227 to 272,229 CFU/ml for TaqMan, 1,824-135,982 for 5x Omni, and 168-6,836 CFU/ml for PerfeCTa. The diagnostic sensitivity using TaqM...
Journal of clinical and diagnostic research : JCDR, 2017
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. To use molecular methods for the improvement of aetiological diagnosis of acute pyogenic meningitis in patients. 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. A total of 39 cases (31.2%) were confir...
Comparison of culture and PCR methods in the diagnosis of bacterial meningitis
Brazilian Journal of Microbiology, 2016
Our aim in this study is to compare the standard culture method with the multiplex PCR and the Speed-Oligo ® Bacterial Meningitis Test (SO-BMT)-a hybridization-based molecular test method-during the CSF examination of the patients with the pre-diagnosis of acute bacterial meningitis. For the purposes of this study, patients with acute bacterial meningitis
Memórias do Instituto Oswaldo Cruz, 2008
Most patients with acute suppurative meningitis are otherwise healthy individuals with regard to immune mechanisms against invasive bacterial disease. This medical emergency is among the most dramatic and potentially ravaging diseases that affect humans, particularly young children. The illness often strikes suddenly, and can either result in death or leave the survivors with significant neurological dysfunctions. The demonstration of a bacterial aetiology is necessary for decisions regarding treatment and prophylaxis. Conventional bacteriological methods frequently fail to identify an agent, as a result of administration of antibiotics or delayed lumbar punctures. We investigated the major aetiologic sources of unspecified bacterial meningitis cases (G00.9, ISCD-10) by polymerase chain reaction (PCR)-based identification of Neisseria meningitidis (crgA), Streptococcus pneumoniae (ply) and Haemophilus influenzae (bexA) in cerebrospinal fluid samples. The multiplex PCR detected N. meningitidis in 92%, S. pneumoniae in 4% and H. influenzae in 1% of the 192 clinical samples assayed; 3% were negative for all three DNA targets. Bacterial DNA detection was found to be a valuable adjunct to enhance bacterial meningitis surveillance when the yield of specimens by culture is reduced. The implementation of PCR assays as a diagnostic procedure in Public Health Laboratories is perceived to be a significant advance in the investigation of bacterial meningitis.
A Multi-Target Real-Time PCR Assay for Rapid Identification of Meningitis-Associated Microorganisms
Molecular Biotechnology, 2013
A central nervous system (CNS) infection, such as meningitis, is a serious and life-threatening condition. Bacterial meningitis can be severe and may result in brain damage, disability or even death. Rapid diagnosis of CNS infections and identification of the pathogenic microorganisms are needed to improve the patient outcome. Bacterial culture of a patient's cerebrospinal fluid (CSF) is currently considered the ''gold standard'' for diagnosing bacterial meningitis. From the CSF cultures researchers can assess the in vitro susceptibility of the causative microorganism to determine the best antibiotic treatment. However, many of the culture assays, such as microscopy and the latex agglutination test are not sensitive. To enhance pathogen detection in CSF samples we developed a multitarget real-time PCR assay that can rapidly identify six different microorganisms: Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, Streptococcus agalactiae, Listeria monocytogenes and Cryptococcus neoformans. In this study we applied this PCR analysis to 296 CSF samples from patients who were suspected of having meningitis. Of the 296 samples that were examined, 59 samples were positive according to the CSF culture and/or molecular assays. Forty-six CSF samples were positive for both the CSF culture and our real-time PCR assay, while 13 samples were positive for the realtime PCR but negative for the traditional assays. This discrepancy may have been caused by the fact that these samples were collected from 23 patients who were treated with antimicrobials before CSF sampling. Keywords Meningitis Á Real-time PCR Á Rapid diagnosis Marco Favaro and Carla Fontana contributed equally to this study.
International Journal of Microbiology
Background. Meningitis is a serious communicable disease with high morbidity and mortality rates. It is an endemic disease in Egypt caused mainly byStreptococcus pneumoniae,Neisseria meningitidis, andHaemophilus influenzae. In some settings, bacterial meningitis is documented depending mainly on positive cerebrospinal fluid (CSF) culture results or CSF positive latex agglutination test, missing the important role of prior antimicrobial intake which can yield negative culture and latex agglutination test results. This study aimed to utilize molecular technology in order to diagnose bacterial meningitis in culture-negative CSF samples.Materials and Methods. Forty culture-negative CSF samples from suspected cases of bacterial meningitis were examined by real-time polymerase chain reaction (real-time PCR) for the presence oflytA,bexA, andctrAgenes specific forStreptococcus pneumoniae,Haemophilus influenzae, andNeisseria meningitidis, respectively.Results. Positive real-time PCR results ...