Preclinical Assessment of a Fully Automated Multiplex PCR Panel for Detection of Central Nervous System Pathogens

Hanson, Kimberly E.; Slechta, E. Susan; Killpack, Jarrett; Heyrend, Caroline; Lunt, Tatum C.; Daly, Judy A.; Hemmert, Andrew; Blaschke, Anne J.

doi:10.1128/jcm.02850-15

Cited by 82 publications

(66 citation statements)

References 14 publications

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“…The study by Leber and colleagues represents the largest clinical assessment of a molecular panel designed to aid in the diagnosis of meningoencephalitis. This study, combined with previous retrospective reports, suggest that ME panel test characteristics are acceptable for clinical care (2,3,7). The main benefit of the ME panel is the potential for more rapid results, which could help inform optimal therapy and resource utilization.…”

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confidence: 74%

“…Most of the viral targets showed excellent agreement with comparator PCRs, even though the multiplex format is slightly less sensitive than the comparator singleplex tests. A notable exception was the calculated sensitivity of the HHV-6 target (86%) and, in a previous report, the cytomegalovirus (CMV) target (57%) (3). Although the ME panel missed two enteroviral infec-tions, the majority of these infections were detected (44/46; sensitivity, 96%).…”

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confidence: 98%

“…The ME panel cannot replace the CSF cryptococcal antigen (CrAg). CrAg testing is fast, cheap, more sensitive, and probably more specific than PCR for the diagnosis of cryptococcal meningitis (3,(5)(6)(7). Whether the ME panel can replace individual viral PCR assays is questionable.…”

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confidence: 99%

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Commentary: The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used?

Hanson

2016

J Clin Microbiol

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Meningitis and encephalitis are potential infectious disease emergencies. The laboratory evaluation of suspected meningitis/encephalitis (ME) is complex, in part because the differential diagnosis is broad and the associated clinical signs and symptoms are not organism specific. Clinicians typically prioritize laboratory workup based on host factors, the duration of symptoms, and potential environmental exposures; but cerebrospinal fluid (CSF) indices in combination with multiple microbiologic tests are generally required to exclude infection or identify a potential pathogen.CSF Gram stain with culture has been the diagnostic "gold standard" for acute bacterial meningitis for nearly a century. However, both methods have a limited sensitivity that may be further reduced in patients who have received antibiotics (1). As a result of these limitations, there has been much interest in the development of standardized molecular diagnostic tests for bacterial central nervous system (CNS) infections. Molecular assays have the potential to be more rapid than culture and are potentially less affected by prior antimicrobial therapy. Until recently, there were no commercially available molecular assays for the diagnosis of bacterial ME marketed in the United States. In contrast, nucleic acid amplification tests are the diagnostic standard for most viral CNS infections. U.S. Food and Drug Administration (FDA)-approved PCR assays have been available for enteroviruses (Xpert EV; Cepheid, Sunnyvale, CA) and herpes simplex viruses (Simplexa HSV 1&2 Direct; Focus Diagnostics, Cypress, CA) for years. Some clinical laboratories have also developed their own PCR assays for CNS pathogens, while others may send testing out to a reference laboratory, which potentially delays time to actionable results.In October 2015, the FDA cleared the first multiplex PCR panel for the detection of CNS pathogens. The fully automated FilmArray ME panel (BioFire Diagnostics LLC, Salt Lake City, UT) simultaneously detects and identifies 14 bacterial, viral, and yeast pathogens in about an hour directly from 200 l of CSF. The components of the FDA-approved FilmArray ME multiplex panel are listed as follows. In this issue of the Journal of Clinical Microbiology, Leber and colleagues report the results of a large multicenter trial designed to support FDA clearance of the ME panel (2). Residual cerebrospinal fluid (CSF) specimens obtained as a part of routine clinical care were tested at 11 different sites in the United States. The ME panel test performance was assessed through comparisons to conventional culture for bacteria or with PCR followed by sequencing for the viral and yeast targets. Discordant results across methods were resolved by repeat molecular analysis (when possible) combined with a blind review of study subject demographic, clinical, and laboratory information.A total of 1,560 prospectively collected CSF specimens were enrolled in the study over 8 months. There was 84.4% positive and Ͼ99.9% negative agreement between the ME panel and conven...

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confidence: 74%

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confidence: 98%

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Commentary: The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used?

Hanson

2016

J Clin Microbiol

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“…There were significant discrepancies between the panel and cryptococcal antigen results, with correlation in only 57% (6/14) of the cryptococcal antigen-positive specimens. Of note, no culture results were available and confirmatory testing by sequencing was also negative (11). Because of the rarity of the infection, L. monocytogenes was not detected in either study; importantly, there were also no false-positive cases identified (10,11).…”

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confidence: 99%

The Brief Case: Neonatal Meningitis Caused by Listeria monocytogenes Diagnosed by Multiplex Molecular Panel

et al. 2016

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c CASE On his 23rd day of life (DOL), a baby boy was transferred to our medical center from a community emergency department for treatment and management of presumed neonatal sepsis and meningitis.He was born full term via normal spontaneous vaginal delivery. His mother was primigravida with no medical complications during her pregnancy. Her prenatal lab results were negative for group B streptococci (GBS) at 36 weeks of gestation, and testing for HIV, syphilis, gonorrhea, chlamydia, and hepatitis B and C was negative. She reported no history of herpes simplex virus (HSV) infection, and no genital lesions were noted on examination. She did not recall having any fever or influenza-like symptoms at any point during her pregnancy. The baby emerged vigorous, was discharged after 2 days in the hospital, was exclusively breastfed, and regained his birth weight by DOL 14.One day prior to his admission on DOL 22, the patient developed a tactile fever, decreased feeding, and fussiness. He was taken to his outpatient pediatrician, who noted an afebrile, well-appearing baby. The baby was given acetaminophen and discharged from the clinic. Over the subsequent night, the patient became fussier and again had tactile fevers. His mother then took him to an emergency department, where he was found to be irritable, tachycardic, and febrile to 39.2°C. No other abnormal examination findings were identified.Blood and cerebrospinal fluid (CSF; obtained by lumbar puncture [LP]) were collected prior to administration of antimicrobials and submitted for culture and laboratory studies (Table 1). His initial CSF was significant for a pleocytosis, as well as abnormally elevated protein (143 mg/dl) and low glucose (16 mg/dl), highly suggestive of bacterial meningitis. The patient was empirically started on intravenous ampicillin (300 mg/kg/day, 4 divided doses), gentamicin (7.5 mg/kg/day, 3 divided doses), and acyclovir (60 mg/kg/day, 3 divided doses) and transferred to our medical center. Upon arrival, he was fussy but afebrile. As the Gram stain was negative for a causative organism, his gentamicin was exchanged for cefotaxime (300 mg/kg/day, 4 divided doses) for more complete empirical Gram-negative coverage and he was continued on ampicillin for empirical treatment of GBS and Listeria, as well as on acyclovir for HSV. Magnetic resonance imaging with contrast of his brain was performed on DOL 24, and mild enhancement of the leptomeninges was noted without any other findings such as abscess or infarction.Blood cultures and CSF cultures remained negative at the community medical center, and an HSV CSF PCR assay was also negative. Repeat blood cultures and CSF were collected on DOL 25. One aerobic blood culture bottle (BD Bactec Peds Plus/F) was collected and incubated in the Bactec automated blood culture system (Becton Dickinson, Franklin Lakes, NJ, USA), and no growth was detected after 5 days. Gram staining of the CSF revealed many white blood cells and no organisms. The specimen was inoculated onto 5% sheep blood agar (blood agar plate...

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“…Lumbar puncture revealed leucocytes 5 × 10 6 /L but no other abnormalities. Cerebrospinal fluid multiplex polymerase chain reaction (PCR) was negative for common bacterial and viral pathogens and specific PCR for herpes simplex viruses 1 and 2, varicella and enterovirus were negative. Serological testing for melioidosis, mycoplasma, hepatitis B and C and human immunodeficiency virus were all negative.…”

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confidence: 99%

Acute myopericarditis in a patient recently started on mesalazine for Crohn disease

Downton

Willcocks

Boutlis

2019

Internal Medicine Journal

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Preclinical Assessment of a Fully Automated Multiplex PCR Panel for Detection of Central Nervous System Pathogens

Cited by 82 publications

References 14 publications

Commentary: The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used?

Commentary: The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used?

The Brief Case: Neonatal Meningitis Caused by Listeria monocytogenes Diagnosed by Multiplex Molecular Panel

Acute myopericarditis in a patient recently started on mesalazine for Crohn disease

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