TABLE 2.

Instruction manuals for detection of phase II IgM antibodies in serum sample^a

Factor or step	Virion/Serion	Biomed	Mikrogen	IBL
Rf absorbent dilution (buffer)	1:4
Serum dilution (buffer)		1:51	1:100	1:100
Serum dilution (Rf absorbent)	1:100	1:1
Rf incubation	RT, 15 min	RT, 30 min
Serum incubation plate	37°C, 1 h	37°C, 1 h	37°C, 1 h	37°C, 1 h
Wash step	4 × 300 μl	4 × 300 μl	3 × 300 μl	3 × 300 μl
Incubation conjugate	37°C, 30 min	37°C, 30 min	RT, 30 min	RT, 30 min
Wash step	4 × 300 μl	4 × 300 μl	3 × 300 μl	3 × 300 μl
Substrate incubation	37°C, 30 min	37°C, 30 min	RT, 15 min, dark	RT, 15 min, dark
Stop	Immediately	Immediately	Immediately	Immediately
Readout (nm)	405/650	405/650	450/620	450/620

^aRT, room temperature.