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A dilution series of the patient's serum is mixed with a suspension of live leptospires in microtiter plates. After incubating for about 2 hr at 30\u00b0C, results are read under the dark\u2010field microscope. The titer is the last dilution in which \u226550% of the leptospires have remained agglutinated. Seroconversion or \u22654\u2010fold titer rise in paired sera is consistent with current leptospirosis. The significance of a titer in a single sample depends on the frequency of residual titers due to past infections and cross\u2010reacting other diseases in the population. Full standardization of the MAT is not possible, but quality assurance can be achieved by participation in the international MAT proficiency testing scheme. <jats:italic>Curr. Protoc. 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