Broad‐Range Bacterial Polymerase Chain Reaction for Early Detection of Bacterial Meningitis

Saravolatz, Louis D.; Manzor, Odette; VanderVelde, Nancy; Pawlak, Joan; Belian, Bradley

doi:10.1086/345438

Cited by 121 publications

(83 citation statements)

References 18 publications

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“…Several studies have demonstrated the usefulness of eubacterial broad-range PCRs for the diagnosis of bacterial meningitis (3,6,13,14,23). However, most published PCR protocols were either time-consuming or did not facilitate species diagnosis of the bacterial pathogens.…”

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confidence: 99%

Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization

et al. 2005

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Real-time PCR and fluorescence in situ hybridization (FISH) were evaluated as rapid methods for the diagnosis of bacterial meningitis and compared to standard diagnostic procedures. For PCR, a LightCycler approach was chosen, implementing eubacterial and specific PCR assays for the most relevant bacteria. For FISH, a similar probe set containing eubacterial and specific probes was composed of published and newly designed probes. Both methods were evaluated by use of cerebrospinal fluid (CSF) samples from patients with suspected bacterial meningitis. For all microscopy-and culture-positive samples (n ‫؍‬ 28), the eubacterial PCR was positive. In addition, all identifiable pathogens were detected with specific PCR assays, according to an algorithm based on the Gram stain. The FISH method detected the pathogen in 13 of 18 positive samples. While the FISH method remained negative for all microscopy-and culture-negative samples (n ‫؍‬ 113), the eubacterial PCR was positive for five of these samples. Sequencing of the amplicon revealed the presence of Neisseria meningitidis, Streptococcus agalactiae, and Haemophilus influenzae in three of these five samples. In addition, samples with discordant results by culture and microscopy were successfully investigated by PCR (10 samples) and FISH (5 samples). In conclusion, PCR is a highly sensitive tool for rapid diagnosis of bacterial meningitis. FISH is less sensitive but is useful for the identification of CSF samples showing bacteria in the Gram stain. Based on our results, an approach for laboratory diagnosis of meningitis including PCR and FISH is discussed.

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confidence: 99%

Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization

et al. 2005

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“…We obtained an acceptable sensitivity of 87.5 % in clinically suspected BM category which was comparable to a study by Rantakokko et al [22] who analyzed CSF samples by PCR technique using 16SrDNA primer and reported a sensitivity of 63.2 %. However, it was less than as in study by Saravoltz et al [23] reported 100 % sensitivity by PCR technique using the same primer. Since, the methodologies have been known separately for eubacterial 16SrDNA as well as for mycobacterial IS6110, our study is first attempt of its kind to give a detailed account of a single tube PCR based detection differentiating eubacterial from mycobacterial in a single PCR run.…”

Section: Discussionmentioning

confidence: 66%

Rapid Diagnosis and Simultaneous Identification of Tuberculous and Bacterial Meningitis by a Newly Developed Duplex Polymerase Chain Reaction

et al. 2015

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The present study describes the development and evaluation of a duplex polymerase chain reaction (D-PCR) for diagnosis and simultaneous identification of tuberculous meningitis (TBM) and bacterial meningitis (BM) in a single reaction. A D-PCR with primers amplifying portions of the Mycobacterium tuberculosis IS6110 and the eubacteria 16SrDNA sequence in a same reaction mix was developed and tested on DNA extracted from 150 clinical CSF samples from different categories (TBM = 39, BM = 26, control infectious and non-infectious category = 85). The results indicate a clear differentiation between bands for eubacteria and M. tuberculosis with an analytical sensitivity of 10 3 cfu/ml for eubacteria and 10 2 cfu/ml for M. tuberculosis. When evaluated in clinical samples, D-PCR overall diagnosed 100 % confirmed TBM and 100 % confirmed BM cases with overall specificity of 96.5 %. D-PCR can be an effective tool for diagnosis and simultaneous identification of TBM or BM in a single PCR reaction. It saves time, cost, labour and sample amount and help in administration of appropriate antimicrobial therapy.The proposed diagnostic assay would be helpful in correct and rapid management of TBM and BM patients.

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“…The application of new diagnostic methods as PCR is substantially increased in routine practice. They can be used as for the detection of bacterial and viral pathogens [8]. It is easy to mistake the early signs and symptoms of Meningococcal disease, especially they are presented as acute respiratory infection, septic shock and lethal outcome within 24 hours.…”

Section: Discussionmentioning

confidence: 99%

Two Cases of Meningococcal Sepsis Caused By Neisseria Meningitidis Serogroup B

Pekova¹

2016

Ann Clin Anal Med

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Thirteen serogroups of bacterium Neisseria meningitidis (N. meningitidis) have been identified, but 5 of them -A, B, C, W-135 and Y -are responsible for majority of the infections worldwide. Serogroup B meningococcus is uncommon in Bulgaria. In 2014 two cases of meningococcal sepsis caused by N. meningitidis serogroup B were diagnosed and treated in the Clinic of Infectious Diseases of University Hospital of Stara Zagora, Bulgaria. They were 10-months old female and 1-year old male. The diagnosis was based on the clinical, epidemiological, laboratorial, microbiological and molecular-genetic analysis. Both cases presented with a sudden oncet of fever, marked asthenia, refusing feeds and hemorrhagic-necrotic skin rash. In the first case the disease evolved to meningococcal sepsis with meningitis with a favorable outcome. The second case rapidly developed fulminant meningococcal sepsis without meningitis with a lethal outcome. In both cases N. meningitidis serogroup B as etiological agent was confirmed.

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Broad‐Range Bacterial Polymerase Chain Reaction for Early Detection of Bacterial Meningitis

Cited by 121 publications

References 18 publications

Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization

Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization

Rapid Diagnosis and Simultaneous Identification of Tuberculous and Bacterial Meningitis by a Newly Developed Duplex Polymerase Chain Reaction

Two Cases of Meningococcal Sepsis Caused By Neisseria Meningitidis Serogroup B

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