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 Introduction of Melioidosis

Melioidosis rapid detection test as shown above picture and it a product of InBios Active Melioidosis DetectTM Rapid Test (AMD) is being developed for the rapid qualitative detection of the capsular polysaccharide produced by the bacterium, Burkholderia pseudomallei. Melioidosis is an infectious disease and its causative agent Burkholderia pseudomallei. This disease is most common in tropical climates, especially in Southeast Asia and northern Australia where it is widespread. Burkholderia pseudomallei is found in contaminated water and soil. It is spread to humans and animals through direct contact with the contaminated source.

Importance of the Melioidosis Rapid Detection Test

• This simple test replaces the use of experienced personnel who have training in principles and the use of microbiological culture identification methods.
• It also replaces the need for a higher biosafety cabinet (BSC) or higher biosafety laboratory (BSL) because of organisms being risk group (risk group 3) except handling bacterial colonies or hemoculture bottles.

Features of This Test Kit

1. It can potentially be used to test whole blood, serum, plasma, urine, pus, sputum, bacterial colonies, or hemoculture bottles.
2.  Results in 15 minutes
3.  Room Temperature Storage
4. Sensitivity of test: 98.7%
5. Specificity: 97.2%

Principle of the Melioidosis Rapid Detection Test

Active Melioidosis DetectTM Rapid Test (AMD) is a lateral flow immunoassay in the form of a nitrocellulose membrane strip coated with a monoclonal antibody (mAB 3C5) to capsular polysaccharide (CPS) produced by Burkholderia pseudomallei during infection. The rapid diagnostic test (RDT) allows detection of the antigen in blood, serum, urine, pus, sputum, and culture fluids. According to the instructions for use, CPS of the closely related Burkholderia mallei is also detected.

Test Requirements

•  The test kit contains RDT strip and buffer( chase) vials
• Sterile test tubes
• Centrifuge
• Biosafety cabinet (class II )
• Blood culture bottle/hemoculture bottles ( specimen)

Test Procedure for Haemoculture Bottle

1. Perform all steps in a Class II biosafety cabinet except for labeling of empty tubes and centrifugation of the samples.
2. Label two sterile tubes for each sample and in one of the tubes,  add two drops of lysis buffer.
3. Centrifuge the cryovial containing 1.5 ml of blood culture broth at 3200×g for 10 minutes.
4. Obtain the supernatants and separated them in the first tube.
5. Suspend the obtained pellet with the lysis buffer in the second tube.
6. Apply 20 μl of this suspension to the sample pad and place strips on paper forms.
7. Add three drops of chase buffer to the inoculated sample pad but here we have used buffer in a tube and left it to soak in the test strip as shown above picture.
8. Set a timer to read the results after 15 minutes.

Observation

• Observe the pink band.
• Test strip: one band at the control and another at the test region

Result and interpretation

1. Only one band at control region: Test Negative
2. Two bands one at control region while another at test region: Test Positive
3. No band at all or band only at test region: Test invalid, in this condition, repeat the test with another test strip.
4. Test: Negative as shown above picture

Bibliography

1. http://inbios.com/wp-content/uploads/2016/06/Melioidosis-Brochure-06.25.14.pdf
2. https://inbios.com/active-melioidosis-detect-rapid-test-kit-intl/
3. https://link.springer.com/article/10.1007/s10096-018-3237-3
4. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4935216/
5. https://wwwnc.cdc.gov/eid/article/21/2/14-1045_article