Beta-Glucan Test

The β-Glucan Test is an in vitro diagnostic test for the quantitative determination of (1→3)-β-D-glucan in serum or plasma. The assay is based on LAL (Limulus Amebocyte lysate) cascade reactions measured by a kinetic turbidimetric method. 

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Test principle

Endotoxin in a sample is inactivated by heating the sample at 70°C for 10 minutes with the pretreatment solution, which contains non-ionic detergent and polymyxin B. This pretreatment also deactivates inhibitory protein substances in the sample. When the pretreated sample is mixed with the LAL reagent, (1→3)-β-D-glucan in the sample activates Factor G which initiates the cascade reactions shown in Fig. 1. The turbidity change caused by the gelation reaction is detected as transmittance change. The time taken for the transmittance to reach the threshold value is measured.  This interval is defined as gelation time (Tg, in Fig. 2) and correlates with (1→3)-β-D-glucan concentration. This concentration is automatically calculated from a standard curve, which is available for each lot of LAL reagent.

Test procedure

1. Pretreatment
0.1 mL of plasma or serum are added to 0.9 mL of pretreatment reagent. After mixing, the sample is heated at 70°C for 10 minutes and cooled on ice subsequently.

Fig. 3: Pretreatment process

2. Measurement
0.2 mL of the pretreated sample is transferred into the LAL reagent vial. After mixing, the sample is inserted into the LIMUSAVE MT-7500 or Toxinometer MT-6500. The measurement starts immediately and takes 90 minutes.

Fig. 4: Measurement process