Detecting Cryptosporidium parvum and Giardia lamblia by coagulation concentration and real-time PCR quantification

• Published in: Frontiers of environmental science & engineering 2013-02, Vol.7 (1), p.49-54
• Main Authors: Zhang, Huining, Zhang, Xiaohu, Zhang, Shuting, Wei, Bo, Jiang, Qipei, Yu, Xin
• Format: Article
• Language: English
• Subjects: Coagulation; Cryptosporidium; Cryptosporidium parvum; Cysts; detection; detection method; DNA; Drinking water; Earth and Environmental Science; Environment; filtration; Giardia; Giardia lamblia; industry; lamblia; method; microbial detection; new methods; Oocysts; parvum; Pathogens; Polymerase chain reaction; Protozoa; quantitative polymerase chain reaction; Real time; Research Article; surface; Surface water; tap water; Turbidity; water; Water supply; Water treatment
• ISSN: 2095-2201; 2095-221X
• DOI: 10.1007/s11783-012-0455-2

Rapid and sensitive pathogen detection methods are essential for the effective functioning of the water treatment industry, yet for many pathogens, effective detection and removal methods are still lacking. Cryptosporidium parvum oocysts and Giardia lamblia cysts are two of the most common waterborne pathogens currently infecting the water supply. In this study, a new method was developed for the detection of Cryptosporidium parvum oocysts and Giardia lamblia cysts. The method includes multi-steps as coagulation concentration of (oo)cysts in water, the dissolution of the resulting flocs into a small volume using acid, filtration of the (oo)cysts solution, and DNA extraction, purification, and examination using real-time PCR. The method was tested using spiked tap water and reservoir water as references, and the mean recovery ranged from 19.6% to 97.6% for oocysts, and from 51.4% to 98.7% for cysts. The method is economical and convenient, and is especially suitable for relatively high turbidity surface water.