Detection and Measurement of Mycoplasma bovis: Correlation of Results using ScanDrop² with CHIPCUVETTE and qTOWER³ Application Note · ScanDrop²

Introduction

Mycoplasma and especially Mycoplasma bovis is a harmful pathogen for cows and calves, which can lead to inflammation and mastitis. Mastitis in turn may result in the reduction of milk production, an important economic factor for farmers in addition to their concern for the health and welfare of their livestock. In veterinary diagnostics it is important to, first, isolate the harmful bacteria from the milk and, second, to detect it correctly. Due to Mycoplasma bovis’ absent cell wall and therefore its natural resistance against antibiotics, therapy is not possible. In essence, spreading of the disease may be stopped only by segregating the infected cows.

As Mycoplasma have no cell well, a special bacteria kit is recommended for the extraction of bacterial nucleic acids from milk. The innuPREP Bacteria DNA Kit from Analytik Jena allows for the isolation of the NA, offering high purity of the DNA and RNA.

Prior to using qualitative real-time PCR to confirm the nature of the extracted NA, it is helpful to check the quality of the samples. This can be done with a relatively simple, fast and inexpensive method using a spectrophotometer. The ScanDrop² from Analytik Jena has been especially designed for these applications, as it gives users the possibility to measure samples spanning a wide range of concentrations.

The CHIPCUVETTE is also particularly well suited for this application, as it allows for the simultaneous measurement of two path lengths for one sample, 0.1 mm and 1 mm. The software can then choose which path length measurement should be used for evaluation, based on a defined set of criteria. Shortly summarized, for samples with concentrations resulting in absorbance values of < 2.0, the 1 mm path length measurement is used. For samples with concentrations resulting in absorbance values >2.0, the 0.1 mm path length measurement is used. This eliminates the need for diluting the sample and saves valuable time in the lab.

For this particular example, samples with high concentrations of extracted bacterial NA may be expected from infected animals. Furthermore, the information concerning the concentration of the target DNA provided by the SanDrop² can help to eliminate false negative results of the qualitative real-time PCR analysis.