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[Establishment of a recombinase-aided isothermal amplification technique to detect Schistosoma japonicum specific gene fragments].

Abstract

OBJECTIVE

To establish a novel method for the detection of Schistosoma japonicum specific gene fragments by recombinase aided isothermal amplification (RAA).

METHODS

The gene fragment SjG28 of S. japonicum was selected as the target gene fragment to be detected, and the primers were designed according to the mechanism of RAA reaction. The reaction of isothermal amplification of S. japonicum was established and optimized. Then this method was applied to amplify and detect the specific gene fragment in the gradient diluent SjG28-recombiant plasmids and different concentrations of S. japonicum genomic DNA to estimate the sensitivity of this method. The samples were also detected by polymerase chain reaction (PCR) in parallel as control. This method was applied to detect the genomic DNA of S. mansoni, Ascaris lumbricoides, and Ancylostoma duodenale to evaluate the specificity.

RESULTS

The specific gene fragment was amplified from genomic DNA of adult worms and eggs of S. japonicum by recombinase aided isothermal amplification reaction established in this study. The reaction can be completed within 30 minutes and the minimum detectable template was 20 copies of plasmids or 0.5 ng of genomic DNA per microliter. Other parasites'genomic DNAs, such as S. mansoni, A. lumbricoides, An. duodenale and healthy human blood genomic DNA were not able to be detected by this method.

CONCLUSIONS

A novel method for the detection of S. japonicum specific gene fragments by recombinase aided isothermal amplification is established in this study, which can be carried out conveniently and rapidly with a considerable sensitivity and specificity, showing the prospect for application in the diagnosis of schistosomiasis japonica.

Authors+Show Affiliations

Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China. Co-first author.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Jiangsu Qitian Gene Technology Co., Ltd., China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.Jiangsu Qitian Gene Technology Co., Ltd., China.Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.

Pub Type(s)

Journal Article

Language

chi

PubMed ID

30019553

Citation

Song, Zhao, et al. "[Establishment of a Recombinase-aided Isothermal Amplification Technique to Detect Schistosoma Japonicum Specific Gene Fragments]." Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi = Chinese Journal of Schistosomiasis Control, vol. 30, no. 3, 2018, pp. 273-277.
Song Z, Ting L, Kun Y, et al. [Establishment of a recombinase-aided isothermal amplification technique to detect Schistosoma japonicum specific gene fragments]. Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2018;30(3):273-277.
Song, Z., Ting, L., Kun, Y., Wei, L., Jian-Feng, Z., Li-Chuan, G., ... Hai-Tao, Y. (2018). [Establishment of a recombinase-aided isothermal amplification technique to detect Schistosoma japonicum specific gene fragments]. Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi = Chinese Journal of Schistosomiasis Control, 30(3), pp. 273-277. doi:10.16250/j.32.1374.2018120.
Song Z, et al. [Establishment of a Recombinase-aided Isothermal Amplification Technique to Detect Schistosoma Japonicum Specific Gene Fragments]. Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2018 Jun 13;30(3):273-277. PubMed PMID: 30019553.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Establishment of a recombinase-aided isothermal amplification technique to detect Schistosoma japonicum specific gene fragments]. AU - Song,Zhao, AU - Ting,Li, AU - Kun,Yang, AU - Wei,Li, AU - Jian-Feng,Zhang, AU - Li-Chuan,Guo, AU - Yan-Hong,Liu, AU - Yang,Dai, AU - Qing-Jie,Ying, AU - Hai-Tao,Yang, PY - 2018/7/19/entrez PY - 2018/7/19/pubmed PY - 2018/7/19/medline KW - Gene fragment KW - Isothermal amplification of nucleic acid KW - Recombinase KW - Schistosoma japonicum SP - 273 EP - 277 JF - Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control JO - Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi VL - 30 IS - 3 N2 - OBJECTIVE: To establish a novel method for the detection of Schistosoma japonicum specific gene fragments by recombinase aided isothermal amplification (RAA). METHODS: The gene fragment SjG28 of S. japonicum was selected as the target gene fragment to be detected, and the primers were designed according to the mechanism of RAA reaction. The reaction of isothermal amplification of S. japonicum was established and optimized. Then this method was applied to amplify and detect the specific gene fragment in the gradient diluent SjG28-recombiant plasmids and different concentrations of S. japonicum genomic DNA to estimate the sensitivity of this method. The samples were also detected by polymerase chain reaction (PCR) in parallel as control. This method was applied to detect the genomic DNA of S. mansoni, Ascaris lumbricoides, and Ancylostoma duodenale to evaluate the specificity. RESULTS: The specific gene fragment was amplified from genomic DNA of adult worms and eggs of S. japonicum by recombinase aided isothermal amplification reaction established in this study. The reaction can be completed within 30 minutes and the minimum detectable template was 20 copies of plasmids or 0.5 ng of genomic DNA per microliter. Other parasites'genomic DNAs, such as S. mansoni, A. lumbricoides, An. duodenale and healthy human blood genomic DNA were not able to be detected by this method. CONCLUSIONS: A novel method for the detection of S. japonicum specific gene fragments by recombinase aided isothermal amplification is established in this study, which can be carried out conveniently and rapidly with a considerable sensitivity and specificity, showing the prospect for application in the diagnosis of schistosomiasis japonica. SN - 1005-6661 UR - https://www.unboundmedicine.com/medline/citation/30019553/[Establishment_of_a_recombinase_aided_isothermal_amplification_technique_to_detect_Schistosoma_japonicum_specific_gene_fragments]_ DB - PRIME DP - Unbound Medicine ER -