Development and evaluation of a thermostatic nucleic acid testing device based on magnesium pyrophosphate precipitation for detecting Enterocytozoon hepatopenaei

Abstract

Enterocytozoon hepatopenaei (EHP) infection has seriously affected prawn culture globally. The symptoms of the infection are not apparent, and traditional detection methods are time consuming and low in accuracy. We developed a new onsite rapid testing device (size 18.8 × 16.7 × 6.6 cm3) for EHP based on magnesium pyrophosphate precipitation and facilitated by loop mediated isothermal amplification (LAMP). The design and fabrication of the device enables efficient light absorbance. The device has a highly sensitive detector, high-precision thermal controller, and humanized touch screen. The temperature control precision of the device is 0.2–0.3 °C at 60 °C, 63 °C, and 65 °C. The coefficients of variation values (CVV) of the luminous power in one channel at light on and off were found to be 0.0097 and 0.0014, respectively, within 1 h. The CVV of the background, luminous power, and values of eight PCR tubes filled with pure water were all less than 5%. In the EHP experiment, eight samples (including seven positive and one negative) confirmed the effectiveness of the device, and four positive and four negative samples verified whether cross-contamination exists. Among them, the rise time of the curve was about 15 min. These results assert that the developed device exhibits enhanced stability and uniformity and has excellent performance with high sensitivity, good specificity, and low testing time. Moreover, the optimal and minimum absorbance range was 555–655 nm for monitoring the production of LAMP.