Detection of Cryptosporidium, Giardia and Toxoplasma in samples of surface water and treated water by multiplex-PCR

Studies on the application of PCR to detect waterborne pathogens have been conducted for more than 15 years but there are still many obstacles in the routine use of molecular biology methods in environmental engineering. The extraction of DNA from environmental materials is one of the most important problem that may occur. While the isolation of DNA from microorganisms found in clean samples is usually successful and the detection by PCR is relatively easy, the variability of the composition of raw water, surface water effectively inhibits PCR. Immunomagnetic Separation (IMS) of cysts/oocysts of protozoa, prior to the DNA extraction phase is the solution which considerably improves the possibility of its detection. Unfortunately, it is possible to implement IMS only against a few pathogens. In the present study the authors demonstrated the possibility of simultaneous detection of the presence of three protozoa in the water without the need of IMS.