scholarly journals Identification of Common Bacterial Pathogens Causing Meningitis in Culture-Negative Cerebrospinal Fluid Samples Using Real-Time Polymerase Chain Reaction

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
Walaa Shawky Khater ◽  
Safia Hamed Elabd
Keyword(s):  
Streptococcus Pneumoniae ◽  
Bacterial Meningitis ◽  
Haemophilus Influenzae ◽  
Neisseria Meningitidis ◽  
Real Time ◽  
Real Time Pcr ◽  
Latex Agglutination ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Culture Negative

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 forStreptococcus pneumoniaewere detected in 36 (90%) of culture-negative CSF samples while no positive results forHaemophilus influenzaeorNeisseria meningitidiswere detected. Four (10%) samples were negative by real-time PCR for all tested organisms.Conclusion. The use of molecular techniques as real-time PCR can provide a valuable addition to the proportion of diagnosed cases of bacterial meningitis especially in settings with high rates of culture-negative results.

scholarly journals Evaluation of Real-Time Polymerase Chain Reaction in Culture-Negative Cerebrospinal Fluid Samples of Bacterial meningitis Patients

2021 ◽  
pp. 96-101
Author(s):  
C. P. Khuntia ◽  
S. K. Kar ◽  
B. Dwibedi
Keyword(s):  
Streptococcus Pneumoniae ◽  
Bacterial Meningitis ◽  
Haemophilus Influenzae ◽  
Real Time ◽  
Real Time Pcr ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Culture Negative

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. Conclusion: The use of molecular techniques as real-time PCR can provide a valuable addition to the proportion of diagnosed cases of bacterial meningitis especially in settings with high rates of culture-negative results.

scholarly journals Detection of Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae in Culture Negative Cerebrospinal Fluid Samples from Meningitis Patients Using a Multiplex Polymerase Chain Reaction in Nepal

2021 ◽  
Vol 13 (1) ◽  
pp. 173-180
Author(s):  
Supriya Sharma ◽  
Jyoti Acharya ◽  
Dominique A. Caugant ◽  
Megha Raj Banjara ◽  
Prakash Ghimire ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cerebrospinal Fluid ◽  
Streptococcus Pneumoniae ◽  
Bacterial Meningitis ◽  
Haemophilus Influenzae ◽  
Neisseria Meningitidis ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Culture Negative

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 statistically not significant (p value > 0.05). In four specimens, culture could not detect any of the targeted bacteria whereas PCR could detect presence of H. influenzae. PCR increases the diagnostic yield for bacterial meningitis. PCR may be considered as an adjunctive test for establishing the cause of infection in culture negative clinically suspected meningitis cases.

scholarly journals Accuracy of real-time PCR, Gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis

2013 ◽  
Vol 13 (1) ◽  
Author(s):  
Henry M Wu ◽  
Soraia M Cordeiro ◽  
Brian H Harcourt ◽  
Maria da Gloria S Carvalho ◽  
Jailton Azevedo ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Haemophilus Influenzae ◽  
Neisseria Meningitidis ◽  
Real Time ◽  
Real Time Pcr ◽  
Gram Stain

scholarly journals Use of a new multiplex quantitative polymerase chain reaction based assay for simultaneous detection of Neisseria meningitidis, Escherichia coli K , Streptococcus agalactiae, and Streptococcus pneumoniae

2021 ◽  
Author(s):  
Nastaran Hemmati ◽  
Farhad Nikkhahi ◽  
Amir Javadi ◽  
Sahar Eskandarion ◽  
Seyed Mahmoud Amin Marashi
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Streptococcus Pneumoniae ◽  
Bacterial Meningitis ◽  
Neisseria Meningitidis ◽  
Streptococcus Agalactiae ◽  
Predictive Value ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain

Background and Objectives: Neisseria meningitidis, Escherichia coli K , Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complica- tions. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. Materials and Methods: The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. Results: The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. Conclusion: The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

scholarly journals Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae and Streptococcus sp. by polymerase chain reaction for the diagnosis of bacterial meningits

2005 ◽  
Vol 63 (4) ◽  
pp. 920-924 ◽  
Author(s):  
Luciane Failace ◽  
Mario Wagner ◽  
Marisa Chesky ◽  
Rosana Scalco ◽  
Luiz Fernando Jobim
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Meningitis ◽  
Haemophilus Influenzae ◽  
Neisseria Meningitidis ◽  
Clinical Status ◽  
Simultaneous Detection ◽  
Etiologic Agent ◽  
Chain Reaction ◽  
Laboratory Findings ◽  
Polymerase Chain

The simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus sp. was assessed by polymerase chain reaction (PCR) for the diagnosis of bacterial meningitis, as well as the applicability of PCR as a routine test. A cohort study was carried out with 182 children (2 months to 12 years of age) with suspicion of bacterial meningitis. Routine tests identified the etiologic agent in 65/84 children whose clinical status and laboratory findings suggested the presence of bacterial meningitis. Bacterial meningitis was ruled out in 98 children. In 19 children, the etiologic diagnosis was not possible using standard methods; in 14 of these patients, the etiologic agent was identified by PCR (N. meningitidis=12; H. influenzae=1; Streptococcus sp.=1). The sensitivity of PCR was 88.1%; specificity, 99.0%; positive predictive value, 98.7%; and negative predictive, 90.1%. PCR is a useful complementary diagnostic technique, especially when Gram stain, culture, or antigenic detection are negative or inconclusive.

scholarly journals DETEKSI CLOSTRIDIUM DIFFICILE TOKSIGENIK MENGGUNAKAN UJI CEPAT TOKSIN DAN REAL TIME POLYMERASE CHAIN REACTION (Toxigenic Clostridium Difficile Detection Using Toxin Rapid Test and Real Time Polymerase Chain Reaction)

2018 ◽  
Vol 22 (1) ◽  
pp. 22
Author(s):  
Ika Yasma Yanti ◽  
Dalima Ari Wahono Astrawinata
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Difficile ◽  
Antibiotic Therapy ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Test ◽  
Chain Reaction ◽  
Chi Square ◽  
Polymerase Chain ◽  
Clostridium Difficile Associated Diarrhea

Toxigenic Clostridium difficile infection, causing a Pseudo Membrane Colitis (PMC) and Clostridium Difficile Associated Diarrhea(CDAD) has increased sharply. The largest risk factor is the use of antibiotics. The purpose of this study was to know how to determinethe prevalence and characteristics of subjects with Toxigenic Clostridium difficile and to assess the ability of the toxin rapid test comparedto real-time PCR. Ninety adult subjects with antibiotic therapy more than two (2) weeks were enrolled in this study. The results of toxinrapid test and real-time PCR were presented in a 2x2 table, statistical test used was Chi square. The prevalence of Toxigenic Clostridiumdifficile based on the toxin rapid test and by real-time PCR was 27.3% and 37.5%, respectively. There were significant differences betweenstool consistency and number of antibiotics used with the detection of Toxigenic Clostridium difficile. There was a relationship betweenthe duration of antibiotic therapy with the detection of Toxigenic Clostridium difficile using real-time PCR (p=0.010, RR=2.116). Thesensitivity, specificity, PPV, NPV, PLR and NLR rapid test against real-time PCR were 69.7%; 98.2%; 95.8%; 84.4%; 39.2 and 0.31,respectively. This study concluded that the prevalence of Clostridium difficile in RSCM was higher compared to that in Malaysia, Thailandand India; the subjects with antibiotic therapy for more than four (4) weeks had a double risk to have Toxigenic Clostridium difficilethan subjects with antibiotic therapy for less than that time (4 weeks). Thus, in this study, toxin rapid test could be used as a tool todetect Toxigenic Clostridium difficile.

Quantitative Real-Time Polymerase Chain Reaction Detection of BK Virus Using Labeled Primers

2010 ◽  
Vol 134 (3) ◽  
pp. 444-448 ◽  
Author(s):  
Zhengming Gu ◽  
Jianmin Pan ◽  
Matthew J. Bankowski ◽  
Randall T. Hayden
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Polymerase Chain Reaction ◽  
Bk Virus ◽  
Clinical Samples ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain

Abstract Context.—BK virus infections among immunocompromised patients are associated with disease of the kidney or urinary bladder. High viral loads, determined by quantitative polymerase chain reaction (PCR), have been correlated with clinical disease. Objective.—To develop and evaluate a novel method for real-time PCR detection and quantification of BK virus using labeled primers. Design.—Patient specimens (n = 54) included 17 plasma, 12 whole blood, and 25 urine samples. DNA was extracted using the MagNA Pure LC Total Nucleic Acid Isolation Kit (Roche Applied Science, Indianapolis, Indiana); sample eluate was PCR-amplified using the labeled primer PCR method. Results were compared with those of a user-developed quantitative real-time PCR method (fluorescence resonance energy transfer probe hybridization). Results.—Labeled primer PCR detected less than 10 copies per reaction and showed quantitative linearity from 101 to 107 copies per reaction. Analytical specificity of labeled primer PCR was 100%. With clinical samples, labeled primer PCR demonstrated a trend toward improved sensitivity compared with the reference method. Quantitative assay comparison showed an R2 value of 0.96 between the 2 assays. Conclusions.—Real-time PCR using labeled primers is highly sensitive and specific for the quantitative detection of BK virus from a variety of clinical specimens. These data demonstrate the applicability of labeled primer PCR for quantitative viral detection and offer a simplified method that removes the need for separate oligonucleotide probes.

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