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Medicine / health science fair project:
Temperature-independent, inexpensive and rapid detection of Ebola




Science Fair Project Information
Title: Temperature-independent, inexpensive and rapid detection of Ebola
Subject: Medicine
Subcategory: Ebola
Grade level: High School - Grades 9-12
Academic Level: Ordinary
Project Type: Building / Engineering
Cost: Medium
Awards: Google Science Fair Finalist
Affiliation: Google Science Fair
Year: 2015
Materials: Silk Fibroin Solution, horseradish peroxidase (HRP)
Techniques: ATR-FTIR analysis, ELISA (Enzyme-Linked Immunosorbent Assay)
Concepts: Ebola Diagnosis
Description: Current detection methods of Ebola are expensive and require uninterrupted refrigeration. This research sought to devise a rapid, simple and inexpensive Ebola detection platform that can be stored and transported without refrigeration. Current Ebola ELISA (Enzyme-Linked Immunosorbent Assay) reagents were embedded in silk fibroin, which possesses stabilizing properties, allowing storage, of otherwise refrigerated reagents, at room temperature. To confirm ELISA colorimetric detection of Ebola after prolonged, non-refrigerated storage of the kit's reagents, the Ebola ELISA was conducted in a 96-wellplate format. Results indicate Ebola ELISA detection is viable in water dilutions only on the day of mixing. For silk-embedded reagents, successful detection was realized for up to one week of storage.
Link: https://www.googlesciencefair.com/projects/en/2015/a035b3
Short Background

Ebola Diagnosis


Electron micrograph of an Ebola virus virion. A micrograph is a photograph or digital image taken through a microscope or similar device to show a magnified image of an item. An electron micrograph is a micrograph prepared using an electron microscope.

Ebola virus disease (EVD) is a viral hemorrhagic fever of humans and other primates caused by ebolaviruses. Signs and symptoms typically start between two days and three weeks after contracting the virus with a fever, sore throat, muscular pain, and headaches. Then, vomiting, diarrhea and rash usually follow, along with decreased function of the liver and kidneys. At this time some people begin to bleed both internally and externally. The disease has a high risk of death, killing between 25 and 90 percent of those infected, with an average of about 50 percent. This is often due to low blood pressure from fluid loss, and typically follows six to sixteen days after symptoms appear.

When EVD is suspected in a person, his or her travel and work history, along with an exposure to wildlife, are important factors to consider with respect to further diagnostic efforts.

Laboratory testing: Possible non-specific laboratory indicators of EVD include a low platelet count; an initially decreased white blood cell count followed by an increased white blood cell count; elevated levels of the liver enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST); and abnormalities in blood clotting often consistent with disseminated intravascular coagulation (DIC) such as a prolonged prothrombin time, partial thromboplastin time, and bleeding time. Filovirions, such as EBOV, may be identified by their unique filamentous shapes in cell cultures examined with electron microscopy, but this method cannot distinguish the various filoviruses.

The specific diagnosis of EVD is confirmed by isolating the virus, detecting its RNA or proteins, or detecting antibodies against the virus in a person's blood. Isolating the virus by cell culture, detecting the viral RNA by polymerase chain reaction (PCR) and detecting proteins by enzyme-linked immunosorbent assay (ELISA) are methods best used in the early stages of the disease and also for detecting the virus in human remains. Detecting antibodies against the virus is most reliable in the later stages of the disease and in those who recover. IgM antibodies are detectable two days after symptom onset and IgG antibodies can be detected 6 to 18 days after symptom onset. During an outbreak, isolation of the virus via cell culture methods is often not feasible. In field or mobile hospitals, the most common and sensitive diagnostic methods are real-time PCR and ELISA. In 2014, with new mobile testing facilities deployed in parts of Liberia, test results were obtained 3–5 hours after sample submission. In 2015 a rapid antigen test which gives results in 15 minutes was approved for use by WHO. It is able to confirm Ebola in 92% of those affected and rule it out in 85% of those not affected.

Early symptoms of EVD may be similar to those of other diseases common in Africa, including malaria and dengue fever. The symptoms are also similar to those of other viral hemorrhagic fevers such as Marburg virus disease.

The complete differential diagnosis is extensive and requires consideration of many other infectious diseases such as typhoid fever, shigellosis, rickettsial diseases, cholera, sepsis, borreliosis, EHEC enteritis, leptospirosis, scrub typhus, plague, Q fever, candidiasis, histoplasmosis, trypanosomiasis, visceral leishmaniasis, measles, and viral hepatitis among others.

Non-infectious diseases that may result in symptoms similar to those of EVD include acute promyelocytic leukemia, hemolytic uremic syndrome, snake envenomation, clotting factor deficiencies/platelet disorders, thrombotic thrombocytopenic purpura, hereditary hemorrhagic telangiectasia, Kawasaki disease, and warfarin poisoning.

Serological tests, if available, are usually the preferred route of identification, however the tests are costly to develop and the reagents used in the test often require refrigeration. Some serological methods are extremely costly, although when commonly used, such as with the "strep test", they can be inexpensive.

See also:
https://en.wikipedia.org/wiki/Ebola_virus_disease
https://en.wikipedia.org/wiki/Ebola_virus
https://en.wikipedia.org/wiki/West_African_Ebola_virus_epidemic

Source: Wikipedia (All text is available under the terms of the GNU Free Documentation License and Creative Commons Attribution-ShareAlike License.)

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