Improved Detection of Bacterial Central Nervous System Infections by Use of a Broad-Range PCR Assay

Meyer, Thomas; Franke, Gefion; Polywka, Susanne; Lütgehetmann, Marc; Gbadamosi, Joystone; Magnus, Tim; Aepfelbacher, Martin

doi:10.1128/jcm.00469-14

Cited by 36 publications

(22 citation statements)

References 18 publications

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“…Many of the pathogens associated with CNS infections are detected by culture, microscopy, or antigen detection techniques. PCR is generally more reliable at detecting pathogens in the CSF, with substantially higher sensitivity than other diagnostic methods such as culture and enzyme-linked immunosorbent assay if the samples are collected at the appropriate time during the infection (16)(17)(18). PCR-based approaches for detecting multiple pathogens in a single array not only increase the number of pathogens that can be detected but also reduce the overall amount of time needed to rule out multiple pathogens.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a TaqMan Array Card for Detection of Central Nervous System Infections

et al. 2017

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Infections of the central nervous system (CNS) are often acute, with significant morbidity and mortality. Routine diagnosis of such infections is limited in developing countries and requires modern equipment in advanced laboratories that may be unavailable to a number of patients in sub-Saharan Africa. We developed a TaqMan array card (TAC) that detects multiple pathogens simultaneously from cerebrospinal fluid. The 21-pathogen CNS multiple-pathogen TAC (CNS-TAC) assay includes two parasites ( and ), six bacterial pathogens (e, ,, ,, and ), and 13 viruses (parechovirus, dengue virus, Nipah virus, varicella-zoster virus, mumps virus, measles virus, lyssavirus, herpes simplex viruses 1 and 2, Epstein-Barr virus, enterovirus, cytomegalovirus, and chikungunya virus). The card also includes human RNase P as a nucleic acid extraction control and an internal manufacturer control, GAPDH (glyceraldehyde-3-phosphate dehydrogenase). This CNS-TAC assay can test up to eight samples for all 21 agents within 2.5 h following nucleic acid extraction. The assay was validated for linearity, limit of detection, sensitivity, and specificity by using either live viruses (dengue, mumps, and measles viruses) or nucleic acid material (Nipah and chikungunya viruses). Of 120 samples tested by individual real-time PCR, 35 were positive for eight different targets, whereas the CNS-TAC assay detected 37 positive samples across nine different targets. The CNS-TAC assays showed 85.6% sensitivity and 96.7% specificity. Therefore, the CNS-TAC assay may be useful for outbreak investigation and surveillance of suspected neurological disease.

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Section: Discussionmentioning

confidence: 99%

Evaluation of a TaqMan Array Card for Detection of Central Nervous System Infections

et al. 2017

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“…A previous study showed 75% sensitivity of conventional CSF culture compared with PCR-based diagnosis. 21 The low sensitivity could be due to the previous use of antimicrobials from the general practitioners that reduced the viable microbes in CSF. In this regard, ME FilmArray platform is a molecular method and empiric use of antimicrobials does not affect its result.…”

Section: Discussionmentioning

confidence: 99%

Childhood Central Nervous System Infections: Are We under Estimating the True Burden?

Jayaweera

Thalagahage²,

Joseph

et al. 2017

J Pediatr Infect Dis

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Children are more susceptible than adults to acquire central nervous system (CNS) infections. This study assesses the validity of routine cerebrospinal fluid (CSF) culture with the meningitis/encephalitis (ME) FilmArray to detect etiological agents in CNS infections. Based on CSF culture, only 5/40 samples were positive in suspected meningitis, whereas 24/40 samples were positive with the ME FilmArray (p?=?0.01). Sensitivity and specificity of conventional culture method were 55.81 and 100%. Overall disability adjusted life years in the study cohort was 1,603.5. Conventional culture with clinical suspicion alone is not enough to obtain an etiological diagnosis of meningitis. The implementation of novel diagnostic techniques is crucial.

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“…Timely access to enterovirus RT-PCR results have been shown to facilitate shorter hospital stays, reduce unnecessary antibiotic use, and lessens ancillary laboratory testing 50– 52 . Broad-range rRNA PCR techniques, which use single pair of primers targeting conserved regions of genes are successfully used for rapid detection and identification of bacterial pathogens and herpesviruses in the CSF 16, 53, 54 . Isothermal amplification-based techniques including loop-mediated isothermal amplification (LAMP) has been successfully developed to provide point care diagnosis within minutes to hours 55 .…”

Section: Molecular Methods In Diagnosis Of Cns Infectionsmentioning

confidence: 99%

“…When compared to conventional methods, molecular methods show greater detection rates: one study reported 16S rRNA PCR-based assays were able to accurately detect the causative organism in 65% of banked CSF samples, compared to 35% when using culture and microscopy 16 . In another report, improved diagnostic yield based on molecular methods were used to optimize antibiotic treatment of patients with infectious meningitis when conventional methods provided a negative result 17 .…”

Section: Introductionmentioning

confidence: 99%

Laboratory Diagnosis of Central Nervous System Infection

Kaplan

Kamboj

et al. 2016

Curr Infect Dis Rep

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CNS infections are potentially life threatening if not diagnosed and treated early. The initial clinical presentations of many CNS infections are nonspecific making a definitive etiologic diagnosis challenging. Nucleic acid in vitro amplification-based molecular methods are increasingly being applied for routine microbial detection. These methods are a vast improvement over conventional techniques with the advantage of rapid turnaround and higher sensitivity and specificity. Additionally, molecular methods performed on cerebrospinal fluid samples are considered the new gold standard for diagnosis of CNS infection caused by pathogens which are otherwise difficult to detect. Commercial diagnostic platforms offer various monoplex and multiplex PCR assays for convenient testing of targets that cause similar clinical illness. Pan-omic molecular platforms possess potential for use in this area. Although molecular methods are predicted to be widely used in diagnosing and monitoring CNS infections, results generated by these methods need to be carefully interpreted in combination with clinical findings. This review summarizes the currently available armamentarium of molecular assays for diagnosis of central nervous system infections, their application, and future approaches.

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Improved Detection of Bacterial Central Nervous System Infections by Use of a Broad-Range PCR Assay

Cited by 36 publications

References 18 publications

Evaluation of a TaqMan Array Card for Detection of Central Nervous System Infections

Evaluation of a TaqMan Array Card for Detection of Central Nervous System Infections

Childhood Central Nervous System Infections: Are We under Estimating the True Burden?

Laboratory Diagnosis of Central Nervous System Infection

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