Abstract
At present, there are few technologies which enable the detection, identification and viability analysis of protozoan pathogens including Cryptosporidium and/or Giardia at the single (oo)cyst level. We report the use of Microfluidic Impedance Cytometry (MIC) to characterise the AC electrical (impedance) properties of single parasites and demonstrate rapid discrimination based on viability and species. Specifically, MIC was used to identify live and inactive C. parvum oocysts with over 90% certainty, whilst also detecting damaged and/or excysted oocysts. Furthermore, discrimination of Cryptosporidium parvum, Cryptosporidium muris and Giardia lamblia, with over 92% certainty was achieved. Enumeration and identification of (oo)cysts can be achieved in a few minutes, which offers a reduction in identification time and labour demands when compared to existing detection methods.
Highlights
The protozoan pathogens Cryptosporidium and Giardia are responsible for a large, global disease burden which affects the health of both humans and livestock and, in turn, has a significant knock-on economic impact[1,2,3]
Utilisation of Microfluidic Impedance Cytometry (MIC) offers for the first time the ability to assess the species and viability status at the singlecyst level, with the additional advantages of rapid automated processing and a label-free, non-destructive method
The impedance properties of C. parvum oocysts of varying source, strain and age were measured in buffers of different conductivity
Summary
The protozoan pathogens Cryptosporidium and Giardia are responsible for a large, global disease burden which affects the health of both humans and livestock and, in turn, has a significant knock-on economic impact[1,2,3]. The standard method of isolation and detection for Cryptosporidium oocysts present in treated water supplies, EPA 1623.117, typically involves the processing of 50–100 L of water (e.g., on site) or 1000 L (e.g., at the treatment works) to a 50 μL volume, before highly-skilled microscopists confirm detection using specialised techniques[18]. This process simultaneously recovers cysts of the Giardia genus – protozoan parasites that may cause gastroenteritis if ingested by humans[17]. There are no reports of the impedance analysis of different Cryptosporidium species, or the simultaneous impedance analysis of Cryptosporidium and Giardia
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