Combining Direct PCR Technology and Capillary Electrophoresis for an Easy-to-Operate and Highly Sensitive Infectious Disease Detection System for Shrimp

Abstract

Infectious diseases are considered the greatest threat to the modern high-density shrimp aquaculture industry. Specificity, rapidity, and sensitivity of molecular diagnostic methods for the detection of asymptomatic infected shrimp allows preventive measures to be taken before disease outbreaks. Routine molecular detection of pathogens in infected shrimp can be made easier with the use of a direct polymerase chain reaction (PCR). In this study, four direct PCR reagent brands were tested, and results showed that the detection signal of direct PCR in hepatopancreatic tissue was more severely affected. In addition, portable capillary electrophoresis was applied to improve sensitivity and specificity, resulting in a pathogen detection limit of 25 copies/PCR-reaction. Juvenile shrimp from five different aquaculture ponds were tested for white spot syndrome virus infection, and the results were consistent with the Organization for Animal Health’s certified standard method. Furthermore, this methodology could be used to examine single post larvae shrimp. The overall detection time was reduced by more than 58.2%. Therefore, the combination of direct PCR and capillary electrophoresis for on-site examination is valuable and has potential as a suitable tool for diagnostic, epidemiological, and pathological studies of shrimp aquaculture.

The Rapid Detection of Salmonella enterica, Listeria monocytogenes, and Staphylococcus aureus via Polymerase Chain Reaction Combined with Magnetic Beads and Capillary Electrophoresis

“…In our present study, we propose an alternative solution for PCR-based detection. Our current PCR-CE system not only offers relative cost-effectiveness compared to other PCR-based techniques but also offers the potential for portability [49]. This feature makes it a valuable tool for on-site pathogen detection.…”

“…A previous study conducted by Lin et al [49] observed that the formulation of PCR reagents obtained from various manufacturers had an impact on amplicon analysis. It is worth noting that Lin et al's study focused on detecting viruses in shrimp samples [49]. They discovered that PCR reagents manufactured by ThermoFisher, Biotools, and BiOptic enabled the detection of the target viruses in their samples, whereas the reagent from Takara (Takara Bio Inc., Tokyo, Japan) did not yield successful results.…”