Graduation Year

2007

Document Type

Thesis

Degree

M.S.P.H.

Degree Granting Department

Public Health

Major Professor

Boo H. Kwa, Ph.D.

Co-Major Professor

Lillian M. Stark, Ph.D.

Committee Member

Donna J. Haiduven, Ph.D.

Keywords

DAPI, EPA, IFA, Method 1622, Method 1623

Abstract

Cryptosporidium parvum is of growing public health concern due to its ability to survive typical water treatment processes. In order to protect the public from infection, the Environmental Protection Agency developed Method 1623 for the detection of Cryptosporidium oocysts in environmental water samples. Execution of this method is time consuming, and the results do not provide an accurate estimation of viability. Therefore, current research is focused on creating a real-time PCR method for the accurate detection of viable Cryptosporidium parvum in environmental water samples.

This thesis presents the development of a real-time PCR method, and the results obtained in its use on field samples. The assay was standardized using multiple dilution series in addition to positive and negative controls. Environmental water samples were tested using this method and Method 1623 for comparison. The results were compared statistically to determine the degree of correlation between methods. The data show that the real-time PCR method correlates well to Method 1623. In addition, the assay was determined to be more cost effective and less labor intensive than Method 1623. Although these early findings are promising, additional research and development are needed before the proposed assay can be used in industry.

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