Rapid Diagnosis Of Bacterial Meningitis Research Articles

The investigation of the efficiency of leukocyte esterase strip test in rapid diagnosis of bacterial meningitis

The investigation of the efficiency of leukocyte esterase strip test in rapid diagnosis of bacterial meningitis

Rapid diagnosis of bacterial meningitis by nanopore 16S amplicon sequencing: A pilot study

Early administration of antibiotics is crucial in the management of bacterial meningitis. Rapid pathogen identification helps to make a definite diagnosis of bacterial meningitis and enables tailored antibiotic treatment. We investigated if the 16S amplicon sequencing performed by MinION, a nanopore sequencer, was capable of rapid pathogen identification in bacterial meningitis. Six retrospective cases of confirmed bacterial meningitis and two prospective cases were included. The initial cerebrospinal fluid (CSF) samples of these patients were used for the experiments. DNA was extracted from the CSF, and PCR was performed on the 16S ribosomal DNA (16S rDNA). Sequencing libraries were prepared using the PCR products, and MinION sequencing was performed for up to 3 h. The reads were aligned to the bacterial database, and the results were compared to the conventional culture studies. Pathogenic bacteria were successfully detected from the CSF by 16S sequencing in all retrospective cases. 16S amplicon sequencing was more sensitive than conventional diagnostic tests and worked properly even in antibiotics-treated samples. MinION sequencing significantly reduced the turnaround time, and even 10 min of sequencing was sufficient for pathogen detection in certain cases. Protocol adjustment could further increase the sensitivity and reduce the turnaround time for MinION sequencing. Finally, the prospective application of MinION 16S sequencing was successful. Nanopore 16S amplicon sequencing is capable of rapid bacterial identification from the CSF of the bacterial meningitis patients. It may have many advantages over conventional diagnostic tests and should therefore be applied in a larger number of patients in the future.

Rapid Diagnosis of Bacterial Meningitis by Urine

Rapid Diagnosis of Bacterial Meningitis by Urine

Enhanced Identification of Group B Streptococcus and Escherichia Coli in Young Infants with Meningitis Using the Biofire Filmarray Meningitis/Encephalitis Panel

FilmArray Meningitis/Encephalitis (ME) polymerase chain reaction (PCR) panel was tested on 62 cerebrospinal fluid (CSF) samples from young infants (0-3 months) with suspected meningitis and compared with CSF cultures. Twelve CSF samples from 9 infants were positive by ME PCR panel (10 Group B Streptococcus (GBS) and 2 Escherichia coli) of which only 5 were positive by culture. The 7 CSF samples that were positive only by ME PCR panel were obtained from infants who had received prior antibiotic treatment. The ME PCR panel can be a useful tool in the rapid diagnosis of bacterial meningitis in pretreated young infants.

Evaluation of high sensitivity C-reactive protein assay in cerebrospinal fluid on the Dimension RxL analyzer

Introduction: Low sensitivity and specificity in traditional laboratory tests became insufficient for accurate diagnostics and initiation of proper treatment of patients infected with bacterial meningitis. High sensitivity C reactive protein (hsCRP) may be an appropriate supplement for rapid diagnosis of bacterial meningitis. The subject of our investigation was the determination of C- reactive protein in cerebrospinal fluid (CSF) duringacute bacterial meningitis.Methods: HsCRP was analysed by a sensitive immunoturbidimetric assay using the Dimension RxL analyser (Siemens). Cerebrospinal fluid concentrations of C-reactive protein have been measured in 20 patients(age range,1 to 50 years) presenting with acute bacterial meningitis and also in a non-infected, non-inflamed control group (n=25).Results: The accuracy and precision of the method proved to be satisfactory. Repeatability of serial sampling for hsCRP described by coefficient of variation were CV=2.1-4.5%. This assay hsCRP in cerebrospinal fluid demonstrates adequate performance characteristics for routine clinical use. Elevated levels of CRP were found in 95% patients with bacterial meningitis. The mean CRP value in 25 uninfected control group was 0.25 mg/L (range 0.10-0.55). The mean CRP for patients with bacterial meningitis was 21.4 mg/L (range 0.40-100).Conclusions: A sensitive assay for CRP in CSF would be an useful adjunct to conventional investigation of acute infective meningitis.

Cerebrospinal Fluid Ferritin for Differentiation of Aseptic and Bacterial Meningitis in Adults

Problem statement: Bacterial meningitis is one of the most important causes of mortality and morbidity. Rapid diagnosis of bacterial cause is very important and critical, because early antibiotic therapy prevents complications; different tests have been developed for rapid diagnosis of bacterial meningitis. One of them is CSF ferritin, which is evaluated in this study. Approach: During cross sectional study from 2008-2009 for 2 years, CSF was collects from 61 adults suspected to have meningitis. These patients admitted to Emam Reza and Sina hospitals of Tabriz. CSF was analyzed for ferritin, glucose, protein, cell count, culture and gram stains. Clinical data's were collected for all patients. Collected data's were analyzed by SPSS software. Results: From 61 specimens, 19 cases had bacterial, 19 cases had aseptic meningitis and 23 cases had no criteria for diagnosis of meningitis. CSF ferritin in bacterial meningitis was 231.63 ± 61.26 ng dL −1 that was significantly higher, than that of aseptic meningitis group. Cut off value for ferritin was estimated 155ng dL −1 with sensitivity of 94% and specificity of 21.7% in this study. Symptoms such as levels of consciousness, fever and seizure were not considered as a criteria for diagnosis of bacterial meningitis (p>0.05). But symptoms such as headache, nausa, vomiting and neck stefness were significantly higher in bacterial and aseptic groups than no meningitis group (p<0.05). Conclusion: In this study CSF ferritin in bacterial meningitis group was significantly higher than aseptic meningitis group. But due to low specificity of CSF ferritin for differentiation of bacterial from aseptic meningitis, we don't recommend performance of this test in early phase of meningitis course.

Survay of Cerebrospinal Fluid (CSF and C-Reactive Protein (CRP) for Differentiation of Bacterial and Viral Meningitis in Loghman Hakim, Aliasghar and Valiasr Hospitals of Zanjan

Background: Bacterial meningitis is a medical emergency which require rapid diagnosis and appropriate therapy. Since the presenting signs and symptoms might be inclusive, laboratory tests are essential in establishing a definite diagnosis. Rapid diagnosis of bacterial meningitis is an important task of most laboratories and is usually based on microbiologic, cytologic and biochemical assays. Measuring serum and CSF C-reactive protein by specially for the countries lacking sophisticated laboratory services. slide latex agglutination is a simple accurate test and is of great value. Material and method: Sixty patients with early symptom of meningitis were hospitalized and subsequently divided in to tow equal groups of bacterial and viral meningitis. CSF and serum samples were collected and required serologic and biochemical testes were performed Results: As compare to viral,bacterial meningitis showed higher increase in CSF routine tests (WBC count, protein) and CPR (P < 0.0001). Glucose showed significant decreased in bacterial meningitis as compare to viral. No significant impact of duration of illness, age, sex and body temperature on CRP value was observed. Conclusion: CRP latex test dose not require expensive equipment or technical expertise and can be performed in any diagnostic laboratory. The simplicity and specificity of this test (sensitivity of 100% in serum and 93.33% in CSF) would encourage its use as a rapid diagnostic test for bacterial meningitis specially in places lacking adequate laboratory facilities.

Rapid diagnosis of bacterial meningitis in children with fluorescence quantitative polymerase chain reaction amplification in the bacterial 16S rRNA gene

Polymerase chain reaction (PCR) techniques have been increasingly used to detect microbial DNA in cerebrospinal fluid (CSF) for the diagnosis of bacterial meningitis. In order to determine the rapidity, sensitivity and specificity of 16S rRNA-based fluorescence quantitative polymerase chain reaction (FQ-PCR), 16S rRNA-based FQ-PCR, CSF bacterial culture and CSF routine analysis were compared in the diagnosis of bacterial meningitis in children. Twenty children who were clinically suspected of bacterial meningitis were included in this study. A total of 2.0 ml of CSF was collected from every child and was subjected to 16S rRNA-based FQ-PCR, CSF culture and CSF routine analysis. Bacterial DNA copies and the cycle threshold (CT) value of the 16S rRNA-based FQ-PCR was recorded, and the results were compared with CSF culture and CSF routine analysis. Seven children were found to be positive with a rate of 35% (7/20) when detected with 16S rRNA-based FQ-PCR and four children displayed a positive rate of 20% (4/20) with the CSF culture method. These two groups displayed a significant difference, with a p-value of 0.002. The method of 16S rRNA-based FQ-PCR demonstrated a high specificity when compared to the standard microbes. A negative correlation was noted between the CT value and the bacteria DNA copies, and the CT value was indicative of the seriousness of bacterial meningitis. 16S rRNA-based FQ-PCR was proved to be a more rapid, sensitive and specific method compared with CSF culture and it should have promising usage in the diagnosis of bacterial meningitis.

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

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.

Rapid Diagnosis of Bacterial Meningitis Using Nitrite Patch Testing

Urine reagent strips have been used to test cerebrospinal fluid (CSF) in areas where laboratory facilities are unavailable. Protein, glucose and leukocyte esterase patches have been shown to be useful in this context. We propose that the nitrite patch also has a contribution to make: it could provide clinically useful information, at no extra cost. We tested CSF samples from 200 children with suspected meningitis. In a pragmatic approach chosen to reflect the clinical dilemma of whether or not to use parenteral antibiotics, the final laboratory diagnosis was dichotomized into either 'bacterial meningitis' or 'not bacterial meningitis'. These diagnostic categories were compared with nitrite patch results, which were either positive or negative. Nitrite patch testing of all CSF, clear CSF and bloody CSF gave positive likelihood ratios of 49, 'infinity' and 5.8, respectively.

Rapid diagnosis of bacterial meningitis by using multiplex PCR and real time PCR

The purpose of the present study was to improve a method for a rapid identification of bacteria in bacterial meningitis by using multiplex polymerase chain reaction (PCR). Ten species of bacteria which cause meningitis in children were investigated, and cerebrospinal fluid from patients with purulent meningitis was studied. The ribosomal RNA genes of bacteria are essential, and are highly conserved in the bacterial kingdoms with consensus region. The 23S rRNA region shows a larger variation among species than in the 16S rRNA region. The authors set primers in the universal region and specific region of 23S rRNA, then amplified these regions by multiplex PCR and real-time PCR. All species of bacteria showed one band by PCR using universal primer. Haemophilus influenzae and Streptococcus pneumoniae showed two bands by multiplex PCR using a combination of universal primers and specific primers. The authors detected H. influenzae within 15 min by using real-time PCR. It was possible to identify clinically significant bacterial species in cerebrospinal fluid by multiplex PCR, and to identify H. influenzae by real-time PCR within a short period.

Rapid diagnosis of bacterial meningitis with reagent strips

Laboratory examination of cerebrospinal fluid (CSF) is not available in many parts of the world, and without knowledge of CSF glucose, protein, and cells, a diagnosis of meningitis may be missed. Testing CSF with urine reagent strips that measure glucose and protein has given variable results. We tested CSF samples from 234 children with suspected meningitis for glucose, protein, and leucocytes with Combur9 reagent strips. The results were compared with those obtained from the laboratory and also interpreted as indicating bacterial or viral meningitis. There was good agreement between the strip and laboratory method of estimating CSF glucose, protein, and leucocytes. All but 4 of the cases of meningitis were correctly identified by the strip method (sensitivity 97%). 2 (2·9%) of 69 cases of bacterial meningitis were judged by an independent observer to be viral, and 2 (3·3%) of 60 cases of viral meningitis as normal. No normal CSF was diagnosed as meningitis (specificity 100%). The results indicate that Combur9 reagent strips can distinguish normal from infected CSF and are of value in the diagnosis of meningitis.

Rapid Diagnosis of Bacterial Meningitis by a Seminested PCR Strategy

A polymerase chain reaction (PCR) based diagnostic assay has been developed for the simultaneous detection in cerebrospinal fluid (CSF) of Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus agalactiae, Listeria monocytogenes and bacteria in general. In the present communication we describe the design of primers for S. pneumoniae, S. agalactiae, and L. monocytogenes, and a general PCR protocol for the assay. The diagnostic outcome is presented for a small collection of CSF specimens including 2 samples from patients with culture-negative purulent meningitis.

Cerebrospinal fluid c-reactive protein measurement: A bedside test in the rapid diagnosis of bacterial meningitis: John M, Raj IS, Macaden R, et al J Trop Pediatr 36: 213–217 Oct 1990

Cerebrospinal fluid c-reactive protein measurement: A bedside test in the rapid diagnosis of bacterial meningitis: John M, Raj IS, Macaden R, et al J Trop Pediatr 36: 213–217 Oct 1990

Cerebrospinal Fluid C-Reactive Protein Measurement--A Bedside Test in the Rapid Diagnosis of Bacterial Meningitis

C-reactive protein (C-RP) determinations were performed by the Latex agglutination method on the cerebrospinal fluid (CSF) samples of 212 patients with clinical features suggestive of meningitis. Patients were grouped as follows Group I: bacterial meningitis and partially treated bacterial meningitis (n = 22). Group II: viral encephalitis (n = 11). Group III: tuberculous meningitis (n = 18). Group IV: (i) febrile convulsions (n = 87); (ii) epileptic seizures (n = 70); (iii) intracranial haemorrhage (n = 4). C-RP was a better indicator of bacterial meningitis (sensitivity 91 per cent) than the Gram's stain (sensitivity 46 per cent). C-RP was positive in 91 per cent of patients in Group I, none in Groups II and III and 0.6 per cent in Group IV. C-RP determination in CSF proved to be a useful indicator of bacterial meningitis and served to distinguish it from viral encephalitis, tuberculous meningitis, febrile convulsions and other central nervous system disorders.

Rapid diagnosis of bacterial meningitis by the detection of a fatty acid marker in CSF with gas chromatography-mass spectrometry and selected ion monitoring

A chemical marker of bacterial meningitis was sought by comparing derivatives of sterile cerebrospinal fluid (CSF) with cultures of organisms in spinal fluid and artificial media. The technique of gas chromatography-mass spectrometry with selected ion monitoring (GC-MS-SIM) was used, optimised for the analysis of fatty acids. Twenty candidate ions were screened, and an ion of mass: charge ratio (m/e) 268 was chosen for detection in clinical specimens. The origin of this marker is unknown, but it is probably the molecular ion of a C16:1 fatty acid. In 135 clinical specimens of CSF examined, the m/e 268 ion was found to be a useful marker for the common organisms that cause bacterial meningitis, giving a sensitivity of 88% and a specificity of 98%. The method was more rapid and more sensitive than conventional microscopy and culture, but CSF containing coagulase-negative staphylococci, Mycobacterium tuberculosis, Cryptococcus neoformans and some other uncommon pathogens gave inconsistent results. Many organisms produced characteristic ion profiles with multiple-ion monitoring, and this method of chemical analysis holds promise for the rapid diagnosis of bacterial infections to genus or species level.

Rapid diagnosis of bacterial meningitis by an enzyme immunoassay of cerebrospinal fluid

A total of 250 cerebrospinal fluid (CSF) specimens were analyzed using a rapid enzyme immunoassay (Pharmacia Meningitis EIA-Test) (EIA) for the detection of antigens of Haemophilus influenzae type b, Neisseria meningitidis (serogroups A,B,C) and Streptococcus pneumoniae (25 selected types). The test is performed in less than 1 h and read by the naked eye. EIA and coagglutination (CoA) were compared with a constructed reference that comprised samples which were either positive by culture and/or on direct microscopy (DM), or in which there were positive results with both EIA and CoA for the bacteria covered by the assays. Using this reference for CSF samples assayed in a period between two meningococcal meningitis epidemics, the sensitivity was 0.86 for EIA and 0.69 for CoA, the specificity 0.95 (EIA) and 0.97 (CoA), the predictive value for a positive result 0.81 (EIA) and 0.87 (CoA) and, the predictive value for a negative result 0.96 (EIA) and 0.93 (CoA). Antibiotics had been given to 54% of the patients before admission. All of the 56 samples that were positive in any of the tests taken during an epidemic of group A meningococcal disease were detected by EIA; CoA was negative in 45% and culture/DM was negative in 32%. Sequential dilutions of two CSF samples from which H. influenzae type b had been isolated, showed the EIA to be 16-32 times more sensitive than CoA. With both technical feasibility and good sensitivity and specificity, the EIA seems to be useful and reliable for the rapid diagnosis of bacterial meningitis, especially in situations where pretreatment with antibiotics are likely.

Cerebral Spinal Fluid

The advent of very sensitive and rapid agglutination tests has been a major advance in facilitating the rapid diagnosis of bacterial meningitis (Table 3). Future investigations will focus on the use of monoclonal antibodies that have been found to be more sensitive and specific than polyclonal antiserums. Modifications of enzyme immunoassays of bacterial antigens to improve speed and technical reliability are in progress. The day may come with the present advances in technology that within 1 hour of receiving a CSF specimen, both the identity and antimicrobial sensitivity of the invading pathogen will be known to the physician. In addition, through quantitation of bacterial antigen, a reliable prognosis as to the outcome of the treated meningitis will also be possible. Finally, the continued refinement of reliable antiserums to group B Neisseria meningitidis, as well as other bacteria responsible for meningitis, particularly in the immunocompromised host will occur.

Rapid diagnosis of bacterial meningitis by antigen detection

affected by prior antimicrobial therapy, which is given liberally in pediatric outpatient settings. Detection of a specific bacterial antigen may be the only means of confirming a suspected diagnosis of bacterial meningitis. The purpose of this article is to review the advantages and disadvantages of the various methods that are commercially available.