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Pre-cut Filter Paper for Detecting Anti-Japanese Encephalitis Virus IgM from Dried Cerebrospinal Fluid Spots
- Source :
- PLoS Neglected Tropical Diseases, PLoS Neglected Tropical Diseases, Vol 10, Iss 3, p e0004516 (2016)
- Publication Year :
- 2016
- Publisher :
- Public Library of Science, 2016.
-
Abstract
- Background The use of filter paper as a simple, inexpensive tool for storage and transportation of blood, ‘Dried Blood Spots’ or Guthrie cards, for diagnostic assays is well-established. In contrast, there are a paucity of diagnostic evaluations of dried cerebrospinal fluid (CSF) spots. These have potential applications in low-resource settings, such as Laos, where laboratory facilities for central nervous system (CNS) diagnostics are only available in Vientiane. In Laos, a major cause of CNS infection is Japanese encephalitis virus (JEV). We aimed to develop a dried CSF spot protocol and to evaluate its diagnostic performance using the World Health Organisation recommended anti-JEV IgM antibody capture enzyme-linked immunosorbent assay (JEV MAC-ELISA). Methodology and Principal Findings Sample volumes, spotting techniques and filter paper type were evaluated using a CSF-substitute of anti-JEV IgM positive serum diluted in Phosphate Buffer Solution (PBS) to end-limits of detection by JEV MAC-ELISA. A conventional protocol, involving eluting one paper punch in 200μl PBS, did not detect the end-dilution, nor did multiple punches utilising diverse spotting techniques. However, pre-cut filter paper enabled saturation with five times the volume of CSF-substitute, sufficiently improving sensitivity to detect the end-dilution. The diagnostic accuracy of this optimised protocol was compared with routine, neat CSF in a pilot, retrospective study of JEV MAC-ELISA on consecutive CSF samples, collected 2009–15, from three Lao hospitals. In comparison to neat CSF, 132 CSF samples stored as dried CSF spots for one month at 25–30°C showed 81.6% (65.7–92.3 95%CI) positive agreement, 96.8% (91.0–99.3 95%CI) negative agreement, with a kappa coefficient of 0.81 (0.70–0.92 95%CI). Conclusions/Significance The novel design of pre-cut filter paper saturated with CSF could provide a useful tool for JEV diagnostics in settings with limited laboratory access. It has the potential to improve national JEV surveillance and inform vaccination policies. The saturation of filter paper has potential use in the wider context of pathogen detection, including dried spots for detecting other analytes in CSF, and other body fluids.<br />Author Summary Japanese encephalitis virus infection is widespread in Asia, and primarily affects children in poor, rural areas. The virus spreads to the brain and spinal cord resulting in significant death and disability. Diagnosis requires testing for the immune response ‘antibody’ specific to Japanese encephalitis virus in the cerebrospinal fluid that surrounds the brain and spinal cord. However, in most areas where the infection occurs there are no laboratory facilities. In the absence of laboratory diagnosis, we have poor knowledge of the extent of the problem in these areas. Dried blood spots on filter paper have been used widely for many years as simple, cost-effective tools for transporting samples for testing. However, there have been few attempts at using dried cerebrospinal fluid spots. We developed a technique for using dried spots to store and test cerebrospinal fluid for antibodies to Japanese encephalitis virus. We compared the results of dried spots with routine neat cerebrospinal fluid in 132 patient samples, and demonstrated excellent agreement between the two tests. This novel method for saturating dried cerebrospinal fluid spots has the potential to enhance our knowledge of Japanese encephalitis virus epidemiology, and inform health policies where they are most needed. It could also be transferred for use in diagnosing other infectious diseases, including using other body fluid samples.
- Subjects :
- RNA viruses
Research Facilities
Physiology
viruses
Pilot Projects
Pathology and Laboratory Medicine
Antibodies, Viral
Nervous System
Geographical Locations
0302 clinical medicine
Cerebrospinal fluid
Filter Paper
Medicine and Health Sciences
030212 general & internal medicine
Enzyme-Linked Immunoassays
Cerebrospinal Fluid
Encephalitis Virus, Japanese
biology
Spots
lcsh:Public aspects of medicine
Temperature
Hematology
3. Good health
Body Fluids
Laboratory Equipment
Infectious Diseases
Blood
Laos
Medical Microbiology
Viral Pathogens
Viruses
Engineering and Technology
Antibody
Anatomy
Pathogens
Research Laboratories
Encephalitis
Research Article
Paper
lcsh:Arctic medicine. Tropical medicine
Asia
lcsh:RC955-962
030231 tropical medicine
Equipment
Research and Analysis Methods
Microbiology
Virus
Mycobacterium
Specimen Handling
03 medical and health sciences
Diagnostic Medicine
medicine
Humans
Desiccation
Immunoassays
Encephalitis, Japanese
Microbial Pathogens
Retrospective Studies
Filter paper
Flaviviruses
business.industry
Public Health, Environmental and Occupational Health
Organisms
Biology and Life Sciences
lcsh:RA1-1270
Japanese encephalitis
medicine.disease
Virology
Immunoglobulin M
Immunology
People and Places
biology.protein
Immunologic Techniques
business
Japanese Encephalitis Virus
Government Laboratories
Subjects
Details
- Language :
- English
- ISSN :
- 19352735 and 19352727
- Volume :
- 10
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....c702b37f56d7842b01b39295a00860c2