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Immunomagnetic separation of Cryptosporidium parvum from source water samples of various turbidities
Z Bukhari1, R M McCuin, C R Fricker
1Clancy Environmental Consultants, St. Albans, Vermont 05478, USA. zbukhari@together.net
Abstract:
Immunomagnetic separation (IMS) procedures which specifically capture Cryptosporidium oocysts and have the potential to isolate oocysts from debris have become commercially available. We compared two IMS kits (kit DB [Dynabeads anti-Cryptosporidium; product no. 730.01; Dynal A.S., Oslo, Norway] and kit IC1 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC, Portland, Maine]) and a modification of kit IC1 (kit IC2 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC]) at three turbidity levels (50, 500, and 5,000 nephelometric turbidity units [ntu]) by using water matrices obtained from different geographical locations. In deionized water, kit DB yielded recoveries between 68 and 83%, whereas the recoveries obtained with kits IC1 and IC2 were more variable and ranged from 0.2 to 74.5%. In water matrices with turbidity levels up to 500 ntu, the oocyst recoveries were more variable with kit DB; however, the recoveries were similar to those obtained in deionized water. In contrast, there were notable reductions in oocyst recoveries in the turbid matrices with kits IC1 and IC2, and the highest recovery (8.3%) was obtained with a 50-ntu sample. An examination of the effects of age on oocyst recovery with kit DB revealed that oocysts up to 16 weeks old yielded recoveries similar to the recoveries observed with fresh oocysts. These data indicate that all IMS kits do not perform equally well, and it is important to conduct in-house quality assurance work before a commercially available IMS kit is selected to replace flotation procedures for recovery of Cryptosporidium oocysts.
Insights
Two immunomagnetic separation (IMS) kits for Cryptosporidium oocyst detection showed variable performance, especially in turbid water. Kit DB demonstrated consistent oocyst recovery across different water conditions and oocyst ages, unlike kits IC1 and IC2.
Area of Science:
- Environmental Science
- Microbiology
- Water Quality Analysis
Background:
- Commercially available immunomagnetic separation (IMS) kits offer potential for isolating Cryptosporidium oocysts from environmental debris.
- Accurate detection of Cryptosporidium oocysts is crucial for public health due to their waterborne transmission and resistance to disinfection.
Purpose of the Study:
- To compare the performance of two IMS kits (DB and IC1) and a modification (IC2) for Cryptosporidium oocyst recovery.
- To evaluate the impact of water turbidity and oocyst age on the efficiency of different IMS kits.
Main Methods:
- Three immunomagnetic separation kits (DB, IC1, IC2) were tested using water matrices with varying turbidity levels (50, 500, 5,000 nephelometric turbidity units [ntu]).
- Oocyst recovery rates were assessed in deionized water and water from different geographical locations.
- The effect of oocyst age (up to 16 weeks) on recovery efficiency was evaluated for one kit (DB).
Main Results:
- Kit DB consistently yielded higher oocyst recoveries (68-83%) in deionized water compared to kits IC1 and IC2 (0.2-74.5%).
- In turbid water (up to 500 ntu), kit DB showed variable but comparable recoveries to deionized water, while kits IC1 and IC2 experienced significant reductions.
- Kit DB demonstrated stable oocyst recovery for oocysts up to 16 weeks old.
Conclusions:
- Immunomagnetic separation kits vary significantly in their efficacy for Cryptosporidium oocyst recovery.
- Water turbidity negatively impacts the performance of certain IMS kits (IC1, IC2), necessitating careful kit selection.
- In-house quality assurance is essential before adopting a commercial IMS kit for routine Cryptosporidium oocyst detection.