Repetitive DNA probes for the detection of Babesia equi.

This research focuses on the development and utilization of DNA probes for identifying Babesia equi, a parasite that affects horses. The team successfully created a genomic library of B. equi and comes up with specific probes that can accurately detect the presence of B. equi DNA.

Construction of Genomic Library

• The researchers initially created a genomic library of Babesia equi. A genomic library is a collection of the total genetic material from a single organism.
• It is made within the bacterium Escherichia coli using a vector called pUC13. A vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell.

Identification of Strongly Hybridizing Clones

• In the next step of the study, they identified several clones that hybridized strongly to B. equi DNA. Hybridization refers to the process of forming a double-stranded DNA molecule between a single-stranded DNA probe and a single-stranded target molecule.
• One particularly successful clone called pBE33 was found to hybridize specifically to B. equi DNA.

Specificity of Clone pBE33

• The team also found that pBE33 did not hybridize with horse DNA. This is significant because it indicates that the clone is highly specific for B. equi and would not confuse the parasite’s genetic material with that of the host organism.
• Additionally, pBE33 also did not hybridize with DNA from other Babesia species such as Babesia caballi, Babesia bovis or Babesia bigemina. This specificity is important for accurate detection and diagnosis.

Development of Repetitive Sequence Probes

• Lastly, the researchers developed two subclones of pBE33 named pSB20 and pEH21, which contain repetitive sequences from B. equi.
• These probes were capable of detecting as little as 0.49ng and 0.97ng of B. equi DNA, respectively, showing high sensitivity and making them useful tools for diagnosing Babesia equi infection in horses.