Table 2.

Comparison of the original and optimized assays using clinical specimens submitted for testing.

Case	Specimens submitted for testing	Original PCR result	Optimized PCR result	Serology results	Case description	Days post symptom onset
Case 1	Seruma	Negative	Ct 37.3	MATb negative	25-year-old male from Ohio, swam in reservoir with case 2, hospitalized in intensive care unit.	4
	Serum	Negative	Negative	MAT negative		5
	Whole blood	Negative	Ct 35.2	Not applicable		6
	Whole blood	Negative	Negative	Not applicable		7
	Whole blood	Negative	Negative	Not applicable		8
	Urine	Negative	Negative	Not applicable		8
Case 2	Whole blood	Negative	Ct 34.3	Not applicable	23-year-old male from Ohio, also swam in reservoir with case 1. Fever and body aches, received fluids.	4, am draw
	Whole blood	Negative	Ct 36.0	Not applicable		4, pm draw
	Serum	Negative	Ct 36.0	MAT negative		4, pm draw
	Whole blood	Negative	Negative	Not applicable		6
	Urine	Negative	Negative	Not applicable		6
	Whole blood	Negative	Negative	Not applicable		7
	Serum	Not tested	Not tested	MAT titer 1:25,600		20
Case 3	CSF	33.5c	35.0	Not applicable	17-year-old male from Iowa with meningitis, swam in river.	7
	Urine	Negative	Negative	Not applicable		9
	Whole blood	Negative	Negative	Not applicable		10
Case 4	Whole blood	Negative	Ct 36.0	Not applicable	23-year-old male from Hawaii with history of water exposure.	5
	Serum	Negative	Negative	MAT titer 1:400		6
	Whole blood	Negative	Negative	Not applicable		18
	Serum	Negative	Negative	MAT titer 1:25,600		18
Case 5	CSF	Negative	34.4	Not applicable	14-year-old with history of travel to Puerto Rico. Positive for leptospirosis using next-generation sequencing (Wilson et al., 2014).	140
	Serum	Negative	Negative	Negatived		190

^aBold text indicates positive results.

^bMicroscopic Agglutination Test.

^cSpecimen was diluted 1:10 due to PCR inhibition.

^dSerology may have been negative due to an immune deficiency.