Ultrasonication of pyrogenic microorganisms improves the detection of pyrogens in the Mono Mac 6 assay.
The monocytic cell line Mono Mac 6 is sensitive to pyrogens. When exposed to pyrogens secretion of interleukin-6 is induced. However, some eukaryotic pyrogenic microorganisms are not detectable. The aim of this study is to introduce a pretreatment of samples to expand the detection range of the assay. The interleukin-6 inducing capacity of a broad spectrum of UV-killed and ultrasonicated microorganisms is examined in Mono Mac 6 cells. The interleukin-6 secretion is determined in a sandwich immunoassay (DELFIA). The Mono Mac 6 assay is able to detect UV-killed Bacillus subtilis, Staphylococcus aureus and Salmonella typhimurium, but neither Candida albicans nor Aspergillus niger. After ultrasonication of the microorganisms it is possible to detect C. albicans and A. niger. The interleukin-6 inducing ability of the examined microorganisms is in no case reduced after ultrasonic treatment. However, ultrasonication of S. aureus results in a 100-fold increase in the interleukin-6 response. Even after ultrasonication Streptococcus faecalis can not be detected. Ultrasonication is an easy and simple method for expanding the detection range in the Mono Mac 6 assay.
The Royal Danish School of Pharmacy, Department of Pharmacology, Denmark. lm@mail.dfh.dk
L
L
Moesby
E W
EW
Hansen
J D
JD
Christensen
D002176:Candida albicans / Q000737:chemistry*; D002460:Cell Line; D006090:Gram-Negative Bacteria / Q000737:chemistry*; D006094:Gram-Positive Bacteria / Q000737:chemistry*; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000382:microbiology*; D011749:Pyrogens / Q000737:chemistry / Q000494:pharmacology; D013010:Sonication
D015850:Interleukin-6; D011749:Pyrogens