Development of a tick-borne pathogen QPCR panel for detection of Anaplasma, Ehrlichia, Rickettsia, and Lyme disease Borrelia in animals.

This research article outlines the development of a test that can detect four of the most common tick-borne diseases in animals such as dogs and horses. These diseases include Anaplasma, Ehrlichia, Rickettsia, and Lyme disease associated Borrelia.

Introduction to Tick-borne Diseases

• Anaplasma, Ehrlichia, Rickettsia, and Lyme disease associated Borrelia are common diseases that ticks can transmit to humans and certain animals like dogs and horses.
• Early detection and accurate diagnosis of these illnesses are important for timely treatment and to avoid serious complications.

The QPCR Panel and Testing Conditions

• The researchers developed a new testing system called the SYBR Green QPCR panel. This system includes four assays that can be used to detect the four mentioned diseases.
• All four assays are optimized to the same thermocycling condition. This condition effectively multiplies the presence of the pathogens’ DNA in a sample to detectable levels.
• The detection limit of these tests is 10 copies per reaction, meaning they can detect the presence of the diseases even if they’re in very low quantities.
• Such sensitivity was confirmed through tests with canine and equine blood samples, spiked with known amounts of synthetic DNA.

Specificity and Accuracy of the QPCR Panel

• The QPCR panel was also found to be specific. This means that it does not react to or detect other pathogens that are often tested in veterinary diagnostic labs. Thus, it avoids false-positive results due to cross reactions.
• With the established Ct cutoff values for positive and negative samples and melting temperature (TM) ranges, the QPCR panel can be used for accurate and rapid screening of the four tick-borne diseases.
• This new method of testing therefore contributes to the accuracy, convenience, and speed of detecting and diagnosing tick-borne diseases in animals.