Molecular Detection and Early Warning of Streptococcus agalactiae and S. iniae
CUI Miao, WU Min, LIU Ru, LI Jingjing, ZHANG Huijie, XU Delin, ZHANG Qizhong
Key Laboratory of Aquatic Eutrophication and Control of Harmful Algal Blooms of Guangdong Higher EducationInstitutes, Engineering Research Center of Tropical and Subtropical Aquatic Ecological Engineering,Ministry of Education, Institute of Hydrobiology, Jinan University, Guangzhou 510632, China
Abstract：Two pairs of specific primers designed according to the 16S rRNA partial gene sequence of Streptococcus agalactiae and the Sip gene sequence of S. iniae, respectively, were applied to establish a rapid and sensitive PCR detection system, aiming to detect trivial amounts of S. agalactiae and S. iniae simultaneously. Two distinct DNA fragments of 870 bp and 614 bp covering the 16S rRNA partial gene sequence and the Sip gene sequence were amplified from the genomic DNA of S. agalactiae and S. iniae, respectively. No cross-reactivity was observed between the two Streptococcus species or with other fish/nonfish pathogens, including Edwards ellatarda, Aeromonasveronii, Vibrio vulnificus, A. sobria, V. alginolyticus, and Escherichia coli. The two primer pairs were capable of detecting genomic DNA of S. agalactiae and S. iniae in concentrations as low as 9.84×10-5 ng/μL and 9.30×10-5 ng/μL, respectively, and of detecting the bacterial cells of S. agalactiae and S. iniae in amount as low as 2.76×103 cfu/mL and 2.51×103 cfu/mL. The duplex PCR designed in this study had strong specificity, high sensitivity, high detection rate, and was used for rapid identification of lower amount of S. agalactiae and S. iniae. The finding provides a valuable tool for early warning detection of the pathogens leading to streptococosis.
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2021, Vol. 40 
