Easy detection of Prorocentrum donghaiense by polymerase chain reaction-nucleic acid chromatography strip

In recent years, Prorocentrum donghaiense, as a dominant species, has ranked first in terms of cumulative number and area of algal blooms in the East China Sea. In this study, the D1–D2 region of the large ribosomal subunit of P. donghaiense was used as the target gene, and specific primers DH–FP/DH–RP were designed according to the species-specific region of the target gene. An easy, sensitive and visual detection method refered to as polymerase chain reaction-nucleic acid chromatography strip (PCR-NACS) was established for P. donghaiense. The optimized parameters of the PCR amplification system are as follows: primer concentration, 0.15 μM; annealing temperature, 62 °C; and Mg²⁺ concentration, 1.5 mM. The specificity test showed that PCR-NACS was exlusively specific for the detection of the target algae. The sensitivity test show that the lowest detection limit (LDL) of PCR-NACS was 2.7 × 10⁻² ng·μL⁻¹ for genomic DNA and 3.58 × 10² copies·μL⁻¹ for plasmid DNA, respectively. The tests using both genomic DNA and plasmid DNA as templates showed that the sensitivity of PCR-NACS was 10 times higher than that of ordinary PCR. The stability test showed that the interfering algal species did not affect the detection of the target algae by PCR-NACS. In addition, the test with simulated natural samples containing target algae showed that the LDL of PCR-NACS could reach 1.27 × 10¹ cells·mL⁻¹. In summary, the PCR-NACS established in this study may provide a new method for easy identification of P. donghaiense in natural water samples.