Performance and Specificity of the Covalently Linked Immunomagnetic Separation-ATP Method for Rapid Detection and Enumeration of Enterococci in Coastal Environments

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2014 Feb 25

PMID 24561583

Citations 2

Authors

Amity G Zimmer-Faust

Vanessa Thulsiraj

Donna Ferguson

Jennifer A Jay

Affiliations

Soon will be listed here.

Abstract

The performance and specificity of the covalently linked immunomagnetic separation-ATP (Cov-IMS/ATP) method for the detection and enumeration of enterococci was evaluated in recreational waters. Cov-IMS/ATP performance was compared with standard methods: defined substrate technology (Enterolert; IDEXX Laboratories), membrane filtration (EPA Method 1600), and an Enterococcus-specific quantitative PCR (qPCR) assay (EPA Method A). We extend previous studies by (i) analyzing the stability of the relationship between the Cov-IMS/ATP method and culture-based methods at different field sites, (ii) evaluating specificity of the assay for seven ATCC Enterococcus species, (iii) identifying cross-reacting organisms binding the antibody-bead complexes with 16S rRNA gene sequencing and evaluating specificity of the assay to five nonenterococcus species, and (iv) conducting preliminary tests of preabsorption as a means of improving the assay. Cov-IMS/ATP was found to perform consistently and with strong agreement rates (based on exceedance/compliance with regulatory limits) of between 83% and 100% compared to the culture-based Enterolert method at a variety of sites with complex inputs. The Cov-IMS/ATP method is specific to five of seven different Enterococcus spp. tested. However, there is potential for nontarget bacteria to bind the antibody, which may be reduced by purification of the IgG serum with preabsorption at problematic sites. The findings of this study help to validate the Cov-IMS/ATP method, suggesting a predictable relationship between the Cov-IMS/ATP method and traditional culture-based methods, which will allow for more widespread application of this rapid and field-portable method for coastal water quality assessment.

Citing Articles

Advancements in mitigating interference in quantitative polymerase chain reaction (qPCR) for microbial water quality monitoring.

Nappier S, Ichida A, Jaglo K, Haugland R, Jones K Sci Total Environ. 2019; 671:732-740.

PMID: 30939326 PMC: 6555561. DOI: 10.1016/j.scitotenv.2019.03.242.

Portable platform for rapid in-field identification of human fecal pollution in water.

Jiang Y, Riedel T, Popoola J, Morrow B, Cai S, Ellington A Water Res. 2017; 131:186-195.

PMID: 29278789 PMC: 5999531. DOI: 10.1016/j.watres.2017.12.023.

References

Caruso G, Monticelli L, Caruso R, Bergamasco A . Development of a fluorescent antibody method for the detection of Enterococcus faecium and its potential for coastal aquatic environment monitoring. Mar Pollut Bull. 2007; 56(2):318-24. DOI: 10.1016/j.marpolbul.2007.10.022. View

Bushon R, Brady A, Likirdopulos C, Cireddu J . Rapid detection of Escherichia coli and enterococci in recreational water using an immunomagnetic separation/adenosine triphosphate technique. J Appl Microbiol. 2009; 106(2):432-41. DOI: 10.1111/j.1365-2672.2008.04011.x. View

Rochelle P, de Leon R, Johnson A, Stewart M, Wolfe R . Evaluation of immunomagnetic separation for recovery of infectious Cryptosporidium parvum oocysts from environmental samples. Appl Environ Microbiol. 1999; 65(2):841-5. PMC: 91105. DOI: 10.1128/AEM.65.2.841-845.1999. View

Hsu B, Huang C . IMS method performance analyses for Giardia in water under differing conditions. Environ Monit Assess. 2006; 131(1-3):129-34. DOI: 10.1007/s10661-006-9462-8. View

Boehm A, Grant S, Kim J, Mowbray S, McGee C, Clark C . Decadal and shorter period variability of surf zone water quality at Huntington Beach, California. Environ Sci Technol. 2002; 36(18):3885-92. DOI: 10.1021/es020524u. View

Lee C, Griffith J, kaiser W, Jay J . Covalently linked immunomagnetic separation/adenosine triphosphate technique (Cov-IMS/ATP) enables rapid, in-field detection and quantification of Escherichia coli and Enterococcus spp. in freshwater and marine environments. J Appl Microbiol. 2010; 109(1):324-33. DOI: 10.1111/j.1365-2672.2009.04660.x. View

Haugland R, Siefring S, Wymer L, Brenner K, Dufour A . Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis. Water Res. 2005; 39(4):559-68. DOI: 10.1016/j.watres.2004.11.011. View

Shanks O, Sivaganesan M, Peed L, Kelty C, Blackwood A, Greene M . Interlaboratory comparison of real-time PCR protocols for quantification of general fecal indicator bacteria. Environ Sci Technol. 2011; 46(2):945-53. DOI: 10.1021/es2031455. View

Field K, Samadpour M . Fecal source tracking, the indicator paradigm, and managing water quality. Water Res. 2007; 41(16):3517-38. DOI: 10.1016/j.watres.2007.06.056. View

10.

Bushon R, Likirdopulos C, Brady A . Comparison of immunomagnetic separation/adenosine triphosphate rapid method to traditional culture-based method for E. coli and enterococci enumeration in wastewater. Water Res. 2009; 43(19):4940-6. DOI: 10.1016/j.watres.2009.06.047. View

11.

Noble R, Griffith J, Blackwood A, Fuhrman J, Gregory J, Hernandez X . Multitiered approach using quantitative PCR to track sources of fecal pollution affecting Santa Monica Bay, California. Appl Environ Microbiol. 2006; 72(2):1604-12. PMC: 1392893. DOI: 10.1128/AEM.72.2.1604-1612.2006. View

12.

Noble R, Blackwood A, Griffith J, McGee C, Weisberg S . Comparison of rapid quantitative PCR-based and conventional culture-based methods for enumeration of Enterococcus spp. and Escherichia coli in recreational waters. Appl Environ Microbiol. 2010; 76(22):7437-43. PMC: 2976187. DOI: 10.1128/AEM.00651-10. View

13.

Saraswat N, Alleman J, Smith T . Enzyme Immunoassay Detection of Nitrosomonas europaea. Appl Environ Microbiol. 1994; 60(6):1969-73. PMC: 201588. DOI: 10.1128/aem.60.6.1969-1973.1994. View

14.

Ferguson D, Moore D, Getrich M, Zhowandai M . Enumeration and speciation of enterococci found in marine and intertidal sediments and coastal water in southern California. J Appl Microbiol. 2005; 99(3):598-608. DOI: 10.1111/j.1365-2672.2005.02660.x. View

15.

Pinto B, PIEROTTI R, Canale G, Reali D . Characterization of 'faecal streptococci' as indicators of faecal pollution and distribution in the environment. Lett Appl Microbiol. 1999; 29(4):258-63. DOI: 10.1046/j.1472-765x.1999.00633.x. View

16.

Ferguson D, Griffith J, McGee C, Weisberg S, Hagedorn C . Comparison of Enterococcus species diversity in marine water and wastewater using Enterolert and EPA Method 1600. J Environ Public Health. 2013; 2013:848049. PMC: 3691910. DOI: 10.1155/2013/848049. View

17.

Ludwig W, Schleifer K . How quantitative is quantitative PCR with respect to cell counts?. Syst Appl Microbiol. 2001; 23(4):556-62. DOI: 10.1016/S0723-2020(00)80030-2. View

18.

Kim J, Grant S . Public mis-notification of coastal water quality: a probabilistic evaluation of posting errors at Huntington Beach, California. Environ Sci Technol. 2004; 38(9):2497-504. DOI: 10.1021/es034382v. View

19.

Maheux A, Dion-Dupont V, Bisson M, Bouchard S, Rodriguez M . Detection of Escherichia coli colonies on confluent plates of chromogenic media used in membrane filtration. J Microbiol Methods. 2013; 97:51-5. DOI: 10.1016/j.mimet.2013.12.008. View

20.

Lee J, Deininger R . Detection of E. coli in beach water within 1 hour using immunomagnetic separation and ATP bioluminescence. Luminescence. 2004; 19(1):31-6. DOI: 10.1002/bio.753. View