Detection of IgM responses to bovine respiratory syncytial virus by indirect ELISA following experimental infection and reinfection of calves: abolition of false positive and false negative results by pre-treatment of sera with protein-G agarose

Abstract

The IgM responses in three panels of sera generated by infection and reinfection of calves with bovine respiratory syncytial virus (BRSV) were measured by indirect ELISA (I-ELISA). The effect of depleting serum IgG by pre-treatment with protein G agarose (PGA) was evaluated. Following primary infection a weak IgM response was detected in the untreated sera of 3 out of 4 calves with maternally derived antibody (MDA). Both the magnitude and duration of the specific IgM responses in these calves were increased by pre-treatment with PGA. In addition, the fourth infected calf tested gave a single positive IgM result following PGA treatment. Transient or persistent IgM responses which were abolished by pre-treatment of sera with PGA were detected in 4/8 calves following reinfection. These were considered to be false positive results, consistent with the influence of IgM rheumatoid factor (IgM-RF). One of these calves and two additional calves showed transient increases in IgM which were resistant to PGA treatment. These were considered to represent specific IgM responses to reinfection. The results indicate the ability of PGA treatment to eliminate both false positive and false negative results and emphasise the necessity for controlling the influence of IgM-RF in IgM-specific indirect ELISAs.

Fig. 1

BRSV-specific IgM (S/P%) and IgG (▴) (COD) responses in Panel 1 sera following primary inoculation of calves with either BRSV strain SVA/274/92 (calves 1–4) or with a control inoculum (calves 5 and 6). BRSV-IgM responses were measured both with (•), and without (□), pre-treatment with PGA. BRSV-IgM PNT = 22% ( – – ).

Fig. 2

BRSV- IgM (S/P%) and -IgG (▴) (COD) responses in Panel 2 sera following reinoculation of calves with either BRSV strain SVA/274/92 (calves 1–4) or with a control inoculum (calves 5 and 6). BRSV-IgM responses were measured both with (•), and without (□), pre-treatment with PGA. BRSV-IgM PNT = 22% ( – – ).

Fig. 3

BRSV-IgM (S/P%) and -IgG (▴) (COD) responses in Panel 3 sera following reinoculation of calves with BRSV strain SVA/187/92. BRSV-IgM responses were measured both with (•), and without (□), pre-treatment with PGA. BRSV-IgM PNT = 22% ( – – ).