New methods for the diagnosis of enteric infections.

Gastrointestinal disease remains a major cause of mortality and morbidity throughout the world. Recently, a number of viral, bacterial, and protozoan agents have been identified which can cause a range of gastrointestinal disorders. The effective management of these diseases requires the prompt identification of the infecting micro-organism and the early institution of preventative and therapeutic interventions. The detection of infecting microorganisms in fecal and intestinal fluids presents a particular challenge to the diagnostic microbiologist. Cultivation can be difficult due to the fastidious nature of the microorganisms and the presence of cytotoxic materials in the specimen. In the past, immunoassays have been used for the detection of some microorganisms. However, immunoassays have limited sensitivity and cannot detect all infecting microorganisms. Recently, nucleic acid amplification techniques have been developed for the direct detection of pathogenic microbial DNA and RNA in human body fluids. We have found that these methods can be applied for the accurate detection of intestinal vlruses provided that inhibitors of enzymatic amplification are removed from the sample. Using affinity binding purification and non-isotopic DNA measurement techniques, we have developed sensitive and specific assays for the quantitation of a wide range of infecting microorganisms in intestinal fluids. Nucleic acid amplification provides a Abstract continued on next page unique tool for the study of enteric pathogens and for the development of strategies for their eventual elimination from the human environment.