Evaluation of multiplex polymerase chain reaction and microarray-based assay for rapid herpesvirus diagnostics

Mannonen, Laura; Vainionpää, Raija; Kauppinen, Juha; Lienhard, Reto; Tritten, Marie-Lise; Cannon, Gemma A.; Hall, William W.; Moilanen, Kirsi; Häkkinen, Maija; Jääskeläinen, Anne J.; Piiparinen, Heli; Mäki, Minna; Järvinen, Anna-Kaarina; Lappalainen, Maija

doi:10.1016/j.diagmicrobio.2012.02.015

Cited by 19 publications

(15 citation statements)

References 42 publications

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“…Enterovirus RNA was detected with RT-PCR, as described previously [19]. DNA for human herpes viruses (varicella zoster (VZV), herpes simplex (HSV) −1 and −2, and human herpesvirus 6 (HHV-6)) was detected with individual qPCRs [20]. The performance of all of the PCR assays has been validated with appropriate proficiency-testing panels from Quality Control for Molecular Diagnostics (Glasgow, Scotland).…”

Section: Methodsmentioning

confidence: 99%

CXCL13 and neopterin concentrations in cerebrospinal fluid of patients with Lyme neuroborreliosis and other diseases that cause neuroinflammation

Hytönen

Kortela

Waris

et al. 2014

J Neuroinflammation

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BackgroundLaboratory diagnosis of Lyme neuroborreliosis (LNB) is partly based on the detection of intrathecal Borrelia burgdorferi–specific antibody production (increased antibody index (AI)). However, AI can be negative in patients with early LNB and, conversely, can remain elevated for months after antibiotic treatment. Recent studies suggested that the chemokine CXCL13 in the cerebrospinal fluid (CSF) is a biomarker for active LNB. Also, CSF neopterin-level determination has been used to assess the degree of neuroinflammation in a wide variety of diseases.MethodsCXCL13 concentrations were analyzed in CSF samples of 366 retrospectively identified individuals. The samples represented pretreatment LNB (38 patients), non-LNB comparison patients, tick-borne encephalitis, central nervous system (CNS) varicella zoster virus infection, CNS herpes simplex virus infection, CNS HHV6 infection, CNS enterovirus infection, and untreated neurosyphilis. The panel included also samples from patients with multiple sclerosis and other neuroinflammatory conditions. Of the LNB patients, 24 posttreatment CSF samples were available for CXCL13 analysis. Neopterin concentrations were determined in a subset of these samples.ResultsThe CXCL13 concentrations in CSF samples of untreated LNB patients were significantly higher (median, 6,480 pg/ml) than the concentrations in the non-LNB group (median, <7.8 pg/ml), viral CNS infection samples (median, <7.8 pg/ml), or samples from patients with noninfectious neuroinflammatory conditions (median, <7.8 pg/ml). The use of cut-off 415 pg/ml led to a sensitivity of 100% and specificity of 99.7% for the diagnosis of LNB in these samples. CSF CXCL13 median concentrations declined significantly from 16,770 pg/ml before to 109 pg/ml after the treatment.CSF neopterin concentration was significantly higher among the untreated LNB patients than in the non-LNB group. The use of neopterin concentration 10.6 nM as the cut-off led to a sensitivity of 88.6% and a specificity of 65.0% for the diagnosis of LNB. The CSF neopterin concentrations decreased statistically significantly with the treatment.ConclusionsThese results clearly indicate that highly elevated CSF CXCL13 levels are strongly associated with untreated LNB. CXCL13 outperformed neopterin and appears to be an excellent biomarker in differentiating LNB from viral CNS infections and from other neuroinflammatory conditions.

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Section: Methodsmentioning

confidence: 99%

CXCL13 and neopterin concentrations in cerebrospinal fluid of patients with Lyme neuroborreliosis and other diseases that cause neuroinflammation

Hytönen

Kortela

Waris

et al. 2014

J Neuroinflammation

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“…Molecular methods for the diagnosis of CNS infections are now well-established in the diagnosis of HSV and enteroviruses. [11121314] Multiplex polymerase chain reaction (PCR) has been successfully used to diagnose viral CNS infections[1516171819202122232425] and bacterial meningitis. [2526]…”

Section: Introductionmentioning

confidence: 99%

Retrospective analysis of multiplex polymerase chain reaction-based molecular diagnostics (SES) in 70 patients with suspected central nervous system infections: A single-center study

Ramalingam¹,

Chakraborty²

2016

Ann Indian Acad Neurol

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Background:Central nervous system (CNS) infections present a grave health care challenge due to high morbidity and mortality. Clinical findings and conventional laboratory assessments are not sufficiently distinct for specific etiologic diagnosis. Identification of pathogens is a key to appropriate therapy.Aim:In this retrospective observational study, we evaluated the efficacy and clinical utility of syndrome evaluation system (SES) for diagnosing clinically suspected CNS infections.Materials and Methods:This retrospective analysis included inpatients in our tertiary level neurointensive care unit (NICU) and ward from February 2010 to December 2013. Cerebrospinal fluid (CSF) samples of 70 patients, clinically suspected of having CNS infections, were subjected to routine laboratory tests, culture, imaging, and SES. We analyzed the efficacy of SES in the diagnosis of CNS infections and its utility in therapeutic decision-making.Results:SES had a clinical sensitivity of 57.4% and clinical specificity of 95.6%. Streptococcus pneumoniae and Pseudomonas aeruginosa were the top two bacterial pathogens, whereas Herpes simplex virus (HSV) was the most common viral pathogen. Polymicrobial infections were detected in 32.14% of SES-positive cases. SES elicited a change in the management in 30% of the patients from initial empiric therapy. At discharge, 51 patients recovered fully while 11 patients had partial recovery. Three-month follow-up showed only six patients to have neurological deficits.Conclusion:In a tertiary care center, etiological microbial diagnosis is central to appropriate therapy and outcomes. Sensitive and accurate multiplex molecular diagnostics play a critical role in not only identifying the causative pathogen but also in helping clinicians to institute appropriate therapy, reduce overuse of antimicrobials, and ensure superior clinical outcomes.

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“…simultaneous detection of HIV and its coinfections (Lochhead et al, 2011), as well as for the diagnostics of atypical pneumonia (Gouriet et al, 2008). As the detection of CNS pathogens, previous microarray designs have focused on the viral nucleic acid detection (Jääskeläinen et al, 2006(Jääskeläinen et al, , 2008Leveque et al, 2011;Mannonen et al, 2012) or detection of herpesvirus, rubella virus and Toxoplasma gondii antibodies using nitrocellulose based or amino-silane activated glass microarrays and proteins as antigens (Bacarese-Hamilton et al, 2004;Jääskeläinen et al, 2009). Proteome microarray has been set up to profile immune response to Borrelia burgdorferi sensu lato (Xu et al, 2008) but no serological or nucleic acid based microarrays have been described earlier for diagnosis of Lyme borreliosis.…”

Section: Introductionmentioning

confidence: 99%

Performance of a multiplexed serological microarray for the detection of antibodies against central nervous system pathogens

Jääskeläinen

Viitala

Kurkela

et al. 2014

Journal of Microbiological Methods

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Central nervous system (CNS) infections have multiple potential causative agents for which simultaneous pathogen screening can provide a useful tool. This study evaluated a multiplexed microarray for the simultaneous detection of antibodies against CNS pathogens. The performance of selected microarray antigens for the detection of IgG antibodies against herpes simplex virus 1 and 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), adenovirus, Mycoplasma pneumoniae and Borrelia burgdorferi sensu lato, was evaluated using serum sample panels tested with reference assays used in a routine diagnostic laboratory. The microarray sensitivity for HSV-1, HSV-2, VZV, adenovirus and M. pneumonia ranged from 77% to 100%, and the specificity ranged from 74% to 97%. Very variable sensitivities and specificities were found for borrelial antigens of three different VlsE protein IR(6) peptide variants (IR6p1, IR6p2, IR6p4) and three recombinant decorin binding proteins A (DbpA; DbpAIa, DbpA91, DbpAG40). For single antigens, good specificity was shown for antigens of IR6p4 and DbpAIa (96%), while DbpA91, IR6p1 and IR6p2 were moderately specific (88-92%). The analytical sensitivity of the microarray was dependent on the borrelial IgG concentration of the specimen. The overall performance and technical features of the platform showed that the platform supports both recombinant proteins, whole viruses and peptides as antigens. This study showed diagnostic potential for all six CNS pathogens, including Borrelia burgdorferi sensu lato, using glutaraldehyde based microarray, and further highlighted the importance of careful antigen selection and the requirement for the use of multiple borrelial antigens in order to increase specificity without a major lack of sensitivity.

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Evaluation of multiplex polymerase chain reaction and microarray-based assay for rapid herpesvirus diagnostics

Cited by 19 publications

References 42 publications

CXCL13 and neopterin concentrations in cerebrospinal fluid of patients with Lyme neuroborreliosis and other diseases that cause neuroinflammation

CXCL13 and neopterin concentrations in cerebrospinal fluid of patients with Lyme neuroborreliosis and other diseases that cause neuroinflammation

Retrospective analysis of multiplex polymerase chain reaction-based molecular diagnostics (SES) in 70 patients with suspected central nervous system infections: A single-center study

Performance of a multiplexed serological microarray for the detection of antibodies against central nervous system pathogens

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