Meticulous Saliva Handling for Alzheimer's Disease Biomarker Analysis
Meticulous Saliva Handling for Alzheimer's Disease Biomarker Analysis

Meticulous Saliva Handling for Alzheimer's Disease Biomarker Analysis

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Saliva has emerged as a promising source of biomarkers for Alzheimer's disease (AD) diagnosis, offering a non-invasive and readily accessible alternative to cerebrospinal fluid (CSF). However, proper collection, purification, and storage of saliva samples are crucial to ensure reliable and accurate biomarker measurements. This article delves into the rigorous protocols employed in the study to maintain the integrity of saliva samples throughout the entire process.

Saliva Sampling:

  1. 1. Participants underwent an 8-hour fasting period and were instructed to refrain from eating, drinking, and smoking for at least 3 hours prior to sample collection to minimize contamination.
  2. 2. Participants rinsed their mouths with water to remove any residual debris or food particles.
  3. 3. Unstimulated whole saliva was collected by asking participants to expectorate into a 15mL polypropylene conical tube, yielding approximately 10mL of saliva.
  4. 4. Immediately after collection, the saliva samples were placed on ice to prevent degradation.

Saliva Purification:

  1. 1. The collected saliva samples were centrifuged at 600g for 10 minutes at 4°C to remove cellular debris and other insoluble materials.
  2. 2. The supernatant was transferred to a new tube, and appropriate protease inhibitors, such as copper sulfate and sodium azide, were added to prevent protein degradation.
  3. 3. Thioflavin S, a dye, was added to prevent amyloid-beta aggregation, and sodium azide, a preservative, was included to inhibit bacterial growth.

Saliva Storage:

  1. 1. The processed saliva samples were aliquoted into 0.5mL microcentrifuge tubes.
  2. 2. The aliquots were stored at -80°C in an ultra-low temperature freezer until further analysis.

The study adhered to stringent standard operating procedures (SOPs) for saliva sample handling to ensure optimal sample quality and minimize potential contamination or degradation. These meticulous steps were critical for preserving the integrity of the low-abundance biomarkers, such as amyloid-beta 42 (Aβ42), present in saliva.

Proper saliva sampling, purification, and storage protocols are essential for reliable biomarker analysis in Alzheimer's disease research. The study's rigorous approach to saliva handling demonstrates the commitment to obtaining high-quality samples, paving the way for accurate and reproducible results. As the field of salivary diagnostics continues to evolve, adherence to such stringent protocols will be paramount in translating promising findings into clinical practice.

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Aβ42 as a Biomarker of Alzheimer’s Disease: Is Saliva a Viable Alternative to Cerebrospinal Fluid?

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