Evaluation of a PCR-immunoassay technique for detection of Neisseria meningitidis in cerebrospinal fluid and peripheral blood

Seward, Rebecca J.; Towner, K. J.

doi:10.1099/0022-1317-49-5-451

Cited by 28 publications

(12 citation statements)

References 7 publications

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“…Since it is more sensitive than culturing, PCR has also been used to investigate clinically suspicious culture-negative cases both in the laboratory and in vaccine trials, increasing the number of confirmed cases of meningococcal disease by up to 60% (24,28,(30)(31)(32). In our study, an additional eight (culturenegative) cases were confirmed by PCR-a 33% increase.…”

Section: Discussionsupporting

confidence: 51%

Prospective Study of a Real-Time PCR That Is Highly Sensitive, Specific, and Clinically Useful for Diagnosis of Meningococcal Disease in Children

Bryant

Zaia

et al. 2004

J Clin Microbiol

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Due to the early administration of antibiotics, meningococcal disease is increasingly difficult to diagnose by culturing. Laboratory studies have shown PCR to be sensitive and specific, but there have been few clinical studies. The objectives of this study were to determine the diagnostic accuracy and clinical usefulness of meningococcal PCR through a prospective comparison of real-time PCR, nested PCR, and standard culturing of blood and cerebrospinal fluid (CSF). The setting was a tertiary-care pediatric hospital in Australia, and the participans were 118 children admitted with possible septicemia or meningitis. The main outcome measuressensitivity, specificity, and positive and negative predictive values-were compared to a "gold standard " fulfilling clinical and laboratory criteria. For 24 cases of meningococcal disease diagnosed by the gold standard, culturing of blood or CSF was positive for 15 (63%), nested PCR was positive for 21 (88%), and real-time PCR was positive for 23 (96%). The sensitivity, specificity, and positive and negative predictive values of real-time PCR (the most sensitive test) for all specimens were, respectively, 96% (95% confidence interval, 79 to 99%), 100% (95% confidence interval, 96 to100%), 100% (95% confidence interval, 85 to 100%), and 99% (95% confidence interval, 94 to 100%). Of 54 patients with suspected meningococcal disease at admission, 23 had positive PCR results. Only one PCR specimen was positive in a patient thought unlikely to have meningococcal disease at admission. Blood PCR remained positive for 33% of patients tested at up to 72 h. Real-time PCR has high positive and negative predictive values in this clinical setting, with better confirmation of cases than nested PCR. Targeting patients for PCR based on admission criteria appears to be practical, and the test may remain useful for several days after the start of antibiotic administration.Meningococcal septicemia and meningitis are life-threatening diseases. Rapid, accurate diagnosis is essential for optimal management of patients and the provision of prompt prophylaxis to contacts. Confirmation of the diagnosis allows physicians to use narrow-spectrum antibiotics, limit the duration of treatment, and provide prognostic information. It also provides vital disease burden information, including data to inform vaccine policy.It is becoming increasingly difficult to confirm the diagnosis of meningococcal infection by conventional microscopy and culturing techniques (6). Blood cultures are positive in about 50% of untreated patients with clinically suspected meningococcal septicemia. This rate is reduced to 5% when antibiotics have been administered prior to admission; primary care practitioners are encouraged in this practice early for suspected cases (8,9,19). Cerebrospinal fluid (CSF) microscopy or culturing is positive in 80 to 90% of untreated cases of meningococcal meningitis, but this rate is also reduced by prior antibiotic administration. In addition, many patients do not undergo a lumbar puncture early in t...

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Section: Discussionsupporting

confidence: 51%

Prospective Study of a Real-Time PCR That Is Highly Sensitive, Specific, and Clinically Useful for Diagnosis of Meningococcal Disease in Children

Bryant

Zaia

et al. 2004

J Clin Microbiol

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“…Adding PCR to the diagnostic tools was a major leap forward in diagnosing systemic meningococcal disease (12,16,17,20,28,32). In the present study, based on the amplification of the capsular transport A gene, we have demonstrated that robotized DNA extraction followed by quantitative PCR is a feasible, rapid, and sensitive diagnostic tool in specifically demonstrating neisserial DNA in patient plasma and CSF.…”

Section: Discussionmentioning

confidence: 84%

“…Quantitative blood culture of meningococci is a cumbersome technique, and few such studies have been published over the years (8,21,30,35). In countries where the health authorities advocate preadmission antibiotic treatment of systemic meningococcal disease, the number of positive blood cultures at hospital admission may now be below 5% (18,28,32).…”

Section: Discussionmentioning

confidence: 99%

Use of Robotized DNA Isolation and Real-Time PCR To Quantify and Identify Close Correlation between Levels of Neisseria meningitidis DNA and Lipopolysaccharides in Plasma and Cerebrospinal Fluid from Patients with Systemic Meningococcal Disease

Øvstebø¹,

Brandtzæg

Brusletto³

et al. 2004

J Clin Microbiol

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The present study, using robotized DNA isolation and quantitative PCR based on the Neisseria meningitidisspecific capsular transport A gene, demonstrates the ease, rapidity, specificity, and sensitivity of quantifying neisserial DNA in plasma (n ‫؍‬ 65) and cerebrospinal fluid (CSF) (n ‫؍‬ 12) from patients with systemic meningococcal disease. We found a close correlation between the levels of neisserial DNA and lipopolysaccharides in plasma (r ‫؍‬ 0.905) and in CSF (r ‫؍‬ 0.964). The median concentration of neisserial DNA in plasma in 23 patients with persistent shock was 2 ؋ 10 7 copies/ml, versus <10 3 copies/ml in 42 nonshock patients. Furthermore, quantitative PCR made possible estimates of the total number of meningococci in plasma, as opposed to conventional blood cultures, suggesting about 1,000 dead meningococci for every viable bacterium. Finally, with logistic regression analyses, neisserial DNA may predict a patient's disease severity and outcome at hospital admission. The number of meningococci in plasma and CSF appears to be the main determinant of the lipopolysaccharide levels, clinical presentation, and outcome.

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“…Permanent sequelae are common among survivors of meningococcal infection (93,95). Definitive diagnosis requires the isolation of N. meningitidis from a normally sterile body fluid or the detection of meningococcal DNA by PCR (36,71,87,259,321).…”

Section: Clinical Aspects Of Meningococcal Infectionmentioning

confidence: 99%

Prospects for Vaccine Prevention of Meningococcal Infection

2006

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SUMMARY Neisseria meningitidis is the leading cause of bacterial meningitis in the United States and worldwide. A serogroup A/C/W-135/Y polysaccharide meningococcal vaccine has been licensed in the United States since 1981 but has not been used universally outside of the military. On 14 January 2005, a polysaccharide conjugate vaccine that covers meningococcal serogroups A, C, W-135, and Y was licensed in the United States for 11- to 55-year-olds and is now recommended for the routine immunization of adolescents and other high-risk groups. This review covers the changing epidemiology of meningococcal disease in the United States, issues related to vaccine prevention, and recommendations on the use of the new vaccine.

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Evaluation of a PCR-immunoassay technique for detection of Neisseria meningitidis in cerebrospinal fluid and peripheral blood

Cited by 28 publications

References 7 publications

Prospective Study of a Real-Time PCR That Is Highly Sensitive, Specific, and Clinically Useful for Diagnosis of Meningococcal Disease in Children

Prospective Study of a Real-Time PCR That Is Highly Sensitive, Specific, and Clinically Useful for Diagnosis of Meningococcal Disease in Children

Use of Robotized DNA Isolation and Real-Time PCR To Quantify and Identify Close Correlation between Levels of Neisseria meningitidis DNA and Lipopolysaccharides in Plasma and Cerebrospinal Fluid from Patients with Systemic Meningococcal Disease

Prospects for Vaccine Prevention of Meningococcal Infection

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