An effective established biosensor of bifunctional probes-labeled AuNPs combined with LAMP for detection of fish pathogen Streptococcus iniae
In purpose of valid Streptococcus iniae detection, we established a colorimetric biosensor using gold nanoparticles (AuNPs) labeled with dual functional probes and along with loop-mediated isothermal amplification (LAMP) assay (LAMP-AuNPs). Based on the characteristics of self-aggregation and bio-conjugation with ligands, AuNPs were chosen for observable color change in tandem with LAMP amplification method to reach high sensitivity and easy operation. Meanwhile, the improvement of dual probes that could fully utilize the LAMP product gave the biosensor a stable result exhibition. LAMP-AuNPs targeting gene ftsB, one of the ATP transporter-related genes, turned out favorable specificity in cross reaction among other fish pathogens. The detect limit of 102 CFU revealed a better sensitivity compared with polymerase chain reaction (PCR) method and AuNPs lateral flow test strip (LFTS). It was also proved to be effective by zebrafish infection model trials with less than 2-h time consumption and nearly no devices which make it a convenient biosensor for point-to-care S. iniae detection.
KeywordsLAMP Gold nanoparticles Bifunctional probes Streptococcus iniae
Conflict of interest
The authors declare that they have no competing interests.
This work was supported by grants from the Ministry of Agriculture of China (nos. CARS-47-G17 and nyhyzx-201303047), the National Natural Sciences Foundation of China (no. 31672696), and the Project of Marine Public Welfare Profession (no. 201405003-2).
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
- Arunrut N, Kampeera J, Sirithammajak S, Sanguanrut P, Proespraiwong P, Suebsing R, Kiatpathomchai W (2016) Sensitive visual detection of AHPND bacteria using loop-mediated isothermal amplification combined with DNA-functionalized gold nanoparticles as probes. PLoS One 11(3):e0151769CrossRefPubMedPubMedCentralGoogle Scholar
- Deng ML, Yu ZH, Geng Y, Wang KY, Chen DF, Huang XL, Ou YP, Chen ZL, Zhong ZJ, Lai WM (2017) Outbreaks of Streptococcosis associated with Streptococcus iniae in Siberian sturgeon (Acipenser baerii) in China. Aquac Res 48(3):909–919Google Scholar
- Dharanivasan G, Riyaz SUM, Jesse DMI, Muthuramalingam TR, Rajendran G, Kathiravan K (2016) DNA templated self-assembly of gold nanoparticle clusters in the colorimetric detection of plant viral DNA using a gold nanoparticle conjugated bifunctional oligonucleotide probe. RSC Adv 6(14):11773–11785CrossRefGoogle Scholar
- Itsaro A, Suanyuk N, Tantikitti C (2012) Multiplex PCR for simultaneous detection of Streptococcus agalactiae, Streptococcus iniae and Lactococcus garvieae: a case of S. agalactiae infection in cultured Nile tilapia (Oreochromis niloticus) and red tilapia (Oreochromis niloticus x Oreochromis moss). Songklanakarin J Sci Technol 34(5):495–500Google Scholar
- Kaewphinit T, Arunrut N, Kiatpathomchai W, Santiwatanakul S, Jaratsing P, Chansiri K (2013) Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples. Biomed Res Int 2013(2013):926230PubMedPubMedCentralGoogle Scholar
- Kundapur RR, Nema V (2016) Loop-mediated isothermal amplification: beyond microbial identification. 1137110Google Scholar
- Shelby R, Shoemaker C, Klesius P (2002) Detection of humoral response to Streptococcus iniae infection of Nile Tilapia, Oreochromis niloticus, by a monoclonal antibody-based ELISA. J Appl Aquac 12(3):23–31Google Scholar
- Storhoff JJ, Lazarides AA, Mucic RC, Mirkin CA, Letsinger RL, Schatz GC (2003) What controls the optical properties of DNA-linked gold nanoparticle assemblies? J Am Chem Soc 125(6):1643–1654Google Scholar
- Wong YP, Othman S, Lau YL, Son R, Chee HY (2017) Loop mediated isothermal amplification (LAMP): a versatile technique for detection of microorganisms. J Appl Microbiol 300(1):83–89Google Scholar
- Zhenyu X, Shaowen K, Chaoqun H, Zhixiong Z, Shifeng W, Yongcan Z (2013) First characterization of bacterial pathogen, Vibrio alginolyticus, for Porites andrewsi white syndrome in the south China sea. PLoS One 8(9):e75425Google Scholar