Establishment of a Nano PCR Assay for Detection of the Streptococcus suis

doi:10.7668/hbnxb.20192014

Abstract

Abstract: This study was to develop an efficient and sensitive nano PCR for the detection of Streptococcus suis (SS). The specific primers were synthesized for the Glutamate dehydrogenase gene (gdh) of SS. Then, the annealing temperature and primer concentration reaction conditions were optimized. The results of specificity assay showed that the method could only amplify 687 bp specific sequence from SS samples, and there was no cross reaction with Actinobacillus pleuropneumoniae (APP), Haemophilus parasuis (H. parasuis), swine Escherichia coli (swine E. coli), Salmonella suis (S. suis) and (S. hyicus). Moreover, the method also could detected the main serotypes 1, 2, 7 and 9 type of SS, respectively. The results of sensitivity assay showed that the lowest SS concentration detected by the nano PCR method was 10 cfu/mL, and its sensitivity was 100 times higher than ordinary PCR of SS. The SS nucleic acid was detected three times, and the results were consistent. Finally, 44 suspected SS infection samples which collected from clinical cases were detected by nano PCR and routine PCR, respectively. The clinical sample testing showed that the positive rates were 72.7% (32/44), 54.5% (24/44), respectively. In conclusion, the nano PCR detection method of SS was established in this study can accurately and efficiently detect SS, which provides technical support for the clinical diagnosis and epidemiological investigation of S. suis disease.

Key words: Streptococcus suis, Glutamate dehydrogenase gene, Nano PCR, Clinical diagnosis

CLC Number:

S855.1

ZHANG Kunli, LIN Benfu, XI Zhenjun, YANG Dongxia, BIAN Zhibiao, SONG Shuai, JIANG Zhiyong, CAI Rujian, LI Chunling. Establishment of a Nano PCR Assay for Detection of the Streptococcus suis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 212-217. doi: 10.7668/hbnxb.20192014.

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