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#16 - Multiplex Assay for Detection of JC Virus
Long Title: Methods and Composition for Identification of Variants of JC Virus DNA for the Diagnostics and Prevention of Progressive Multifocal Leukoencephalopathy (PML)
NIH Reference No.: E-088-2012
Executive Summary
General Description
The JC virus infects an estimated 70 percent of people worldwide, but it is usually harmless due to a healthy immune response. The virus, however, is never cleared entirely from the body; instead it remains inactive inside hidden reservoirs. When the body is in an immuno-compromised state, such as in the case of cancer, HIV, or multiple sclerosis, JC virus can sometimes reactivate, resulting in Progressive Multifocal Leukoencephalopathy (PML). PML can prove fatal within months or even weeks. In recent years, several cases of PML have been reported in individuals receiving certain types of monoclonal antibody therapies (mAb), cutting-edge medications that can successfully treat multiple sclerosis (MS), Crohn’s disease, rheumatoid arthritis and other autoimmune disorders. PML is an orphan indication and occurs in approximately one in 200,000 people and has a broad geographical distribution. In general, PML has a mortality rate of 30-50 percent in the first few months following diagnosis but depends on the severity of the underlying disease and treatment received. There is no known treatment for PML. Brain biopsy remains the gold standard confirmatory test for diagnosis of PML.
The invention describes JCV qPCR multiplex assay that targets two regions in JCV genomes to simultaneously identify and measure viral DNA, as well as distinguish between virulent (causes PML) and non-virulent JCV (does not cause PML). The results can signal risk for PML if patients test positive for JCV variants closely associated with PML pathogenesis. The results can be extrapolated to help clinicians determine which patients are good candidates for mAb therapies and when a patient may be at risk of developing PML from the mAb therapy during a course of treatment.
Scientific Progress
Testing was performed using control plasmids and patients' cerebrospinal fluid (CSF), blood, and urine from PML patients. It predictably demonstrated the non-virulent, archetype non-coding regulatory region (NCRR) was present in urine, but virulent NCRR variants were present in CSF and blood. Thus, a single assay quantifies and identifies JCV DNA in clinical samples and discriminates between virulent/non-virulent variants.
Future Work
Strengths
Weakness
Patent Status
PCT Application No. PCT/US2013/046158 filed June 17, 2013
Publications
Ryschkewitsch CF, et al., J Clin Virol. 2013. 57(3):243-8 (PMID: 23619054)
Berger JR, et al., Neurology 2013. 80(15):1430-8 (PMID: 23568998)
Inventor Bio
Eugene Major Ph.D.
Eugene O. "Gene" Major, PhD, is a senior investigator at the National Institute of Neurological Disorders and Stroke (NINDS) of NIH. Dr. Major conducts research into the neurological diseases including progressive multifocal leukoencephalopathy (PML), caused by JC virus and often found in immunosuppressed patients such as those with HIV/AIDS. Dr. Major has published over 140 scientific articles and reviews in the peer-reviewed literature and has contributed to Fields Virology, a standard virology textbook. Dr. Major has been quoted extensively in news coverage of the finding that natalizumab (Tysabri) and related monoclonal antibody-based therapies increase the risk of a rare brain disease caused by JC virus.
NIH Reference No.: E-088-2012
Executive Summary
- Invention Type: Diagnostics
- Patent status: PCT application (PCT/US2013/046158) filed June 17, 2013
- LINK: http://www.ott.nih.gov/technology/e-088-20120
- NIH Reference Number: E-088-2012
- NIH Institute or Center: National Institute of Neurological Disorders and Stroke (NINDS)
- Disease focus: JC Virus infection, progressive multifocal leukoencephalopathy
- Basis of Invention: Multiplex Assay
- How it works: Identification of the JC virus, based on the presence of unique viral nucleic acid sequence
- Lead Inventors: Eugene Major (NINDS); Caroline Ryschkewitsch (NINDS)
- Development Stage: Currently in use under CLIA certification
- Novelty: High sensitivity of detection, No need for sequencing
- Clinical Application: The method can be developed into a diagnostic kit
General Description
The JC virus infects an estimated 70 percent of people worldwide, but it is usually harmless due to a healthy immune response. The virus, however, is never cleared entirely from the body; instead it remains inactive inside hidden reservoirs. When the body is in an immuno-compromised state, such as in the case of cancer, HIV, or multiple sclerosis, JC virus can sometimes reactivate, resulting in Progressive Multifocal Leukoencephalopathy (PML). PML can prove fatal within months or even weeks. In recent years, several cases of PML have been reported in individuals receiving certain types of monoclonal antibody therapies (mAb), cutting-edge medications that can successfully treat multiple sclerosis (MS), Crohn’s disease, rheumatoid arthritis and other autoimmune disorders. PML is an orphan indication and occurs in approximately one in 200,000 people and has a broad geographical distribution. In general, PML has a mortality rate of 30-50 percent in the first few months following diagnosis but depends on the severity of the underlying disease and treatment received. There is no known treatment for PML. Brain biopsy remains the gold standard confirmatory test for diagnosis of PML.
The invention describes JCV qPCR multiplex assay that targets two regions in JCV genomes to simultaneously identify and measure viral DNA, as well as distinguish between virulent (causes PML) and non-virulent JCV (does not cause PML). The results can signal risk for PML if patients test positive for JCV variants closely associated with PML pathogenesis. The results can be extrapolated to help clinicians determine which patients are good candidates for mAb therapies and when a patient may be at risk of developing PML from the mAb therapy during a course of treatment.
Scientific Progress
Testing was performed using control plasmids and patients' cerebrospinal fluid (CSF), blood, and urine from PML patients. It predictably demonstrated the non-virulent, archetype non-coding regulatory region (NCRR) was present in urine, but virulent NCRR variants were present in CSF and blood. Thus, a single assay quantifies and identifies JCV DNA in clinical samples and discriminates between virulent/non-virulent variants.
Future Work
- The original work was done using 52 PML patients and 27 non-PML patients. Larger cohort needed for further analysis of the false-positive/negative rates.
- Testing urine and/or plasma samples over time from patients in risk categories to monitor disease progression
Strengths
- Orphan indication
- DNA sequencing not required
- Single assay format using same template to identify prototype and archetype with 10c/ml sensitivity
- Large market for clinical diagnostics to monitor effect of mAb on levels of JC Virus
Weakness
- Reliance on JC virus PCR alone to demonstrate the presence of the virus in tissue remains investigational
- JC virus viremia can occur in healthy individuals; any contamination of the CSF with blood has the potential for providing a false-positive result
- False Positives: Although amplification of the virus from the CSF in the absence of PML has been considered very unlikely, a low copy number of JC virus was reported in 2 of 515 CSF samples from patients without PML using these ultrasensitive techniques
- False Negatives: Despite the high sensitivity of the PCR assay, a negative PCR does not rule out PML, and in some cases biopsy of the brain with PCR amplification from the brain tissue has been employed to establish the diagnosis
- Even if PML diagnosis occurs, there is no effective treatment at present. There is one in vitro compound that seems promising; CMX001 (see treatment section here: http://aidsinfo.nih.gov/guidelines/html/5/pediatric-oi-prevention-and-treatment-guidelines/416/progressive-multifocal-leukoencephalopathy)
Patent Status
PCT Application No. PCT/US2013/046158 filed June 17, 2013
Publications
Ryschkewitsch CF, et al., J Clin Virol. 2013. 57(3):243-8 (PMID: 23619054)
Berger JR, et al., Neurology 2013. 80(15):1430-8 (PMID: 23568998)
Inventor Bio
Eugene Major Ph.D.
Eugene O. "Gene" Major, PhD, is a senior investigator at the National Institute of Neurological Disorders and Stroke (NINDS) of NIH. Dr. Major conducts research into the neurological diseases including progressive multifocal leukoencephalopathy (PML), caused by JC virus and often found in immunosuppressed patients such as those with HIV/AIDS. Dr. Major has published over 140 scientific articles and reviews in the peer-reviewed literature and has contributed to Fields Virology, a standard virology textbook. Dr. Major has been quoted extensively in news coverage of the finding that natalizumab (Tysabri) and related monoclonal antibody-based therapies increase the risk of a rare brain disease caused by JC virus.