SepsiTest™-UMD Add-on 10 Kit

The SepsiTest-UMD Add-on 10 Kit is a pre-treatment for larger blood volumes, used ahead of SepsiTest-UMD or UMD-SelectNA. In sepsis the bacterial load in blood is often only a few colony forming units per millilitre, so the volume examined determines what can be found.

Principle

Buffer CM lyses the human cells in the sample and the released human DNA is degraded enzymatically by MolDNase B. The pathogen cells and cell debris are then separated by centrifugation at 9,500 x g for 10 minutes and taken into the DNA isolation of the downstream kit.

Sensitivity

With a sample volume of 10 ml the manufacturer reports: Staphylococcus aureus strain 1 below 6 cfu/ml and strain 2 at 1 cfu/ml, methicillin-resistant S. aureus below 5 cfu/ml, Escherichia coli 7 cfu/ml and Candida albicans 1 cfu/ml.

Sample material and limitations

Whole blood and other primary sterile body fluids, 1 to 10 ml, with a protocol for more than 1 to 5 ml and one for more than 5 to 10 ml. The CE marking covers the detection of bacteria and fungi without further taxonomic differentiation. The manufacturer states that the test cannot serve as the sole diagnostic tool, that positive results require confirmation by sequencing, and that skin flora contaminating the sample may be detected. Storage is at room temperature, in the dark, and not refrigerated.

1. Sepsitest UMD PCR Master Mix: A solution containing all of the reagents necessary for PCR amplification of the captured microbial DNA from the Sepsitest UMD kit.
2. Control DNA: A synthetic DNA that is added to the sample as an internal control to ensure that the assay is working correctly.
3. PCR Primers: Specific primers that amplify conserved regions of the microbial DNA for detection of additional pathogens.
4. PCR Enzyme: A heat-stable DNA polymerase enzyme used to catalyze the PCR reaction.
5. DNase/RNase-free Water: Water that has been treated to remove contaminating DNases and RNases, which can degrade the extracted DNA.

The kit does not include the magnetic beads or lysis buffer that are included in the original Sepsitest UMD kit, as these are assumed to have already been used for the initial detection of microbial DNA in the blood sample. The Sepsitest UMD Add-On 10 kit is designed to be used in conjunction with the original Sepsitest UMD kit for the detection of additional pathogens.