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Establishment of LAMP Assay for Detection of Bacillus cereus

Establishment of LAMP Assay for Detection of Bacillus cereus

Xu Yun-Ming1*, Sun Zhi-Yuan1, Yang Jian-Bo1, Bian Rong-Rong2, Ren Hong-Lin3, Zhong Si-Yuan1, Cai Yu-Hong1, Peng Jing1 and Bao Hua-Xia1

1Institute of Animal Husbandry and Veterinary Medicine, Jiangsu Vocational College of Agriculture and Forestry, Jurong 212400, China
2Bureau of Agricultural and Rural Affairs of Jurong, Jurong 212400, China; 
3Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, Jilin University. Changchun 130062, China
 
* Corresponding author: xym2013@jsafc.edu.cn

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Fig. 1.

Detection specificity of LAMP reaction for strains. M, DL2000 DNA marker; N, negative control. 1-2, Bacillus cereus; 3, Yersinia enterocolitica; 4, Vibrio prholyticus; 5, Candida albicans; 6, Salmonella choleraesuls; 7, Corynebacterium glutamicum; 8, Staphylococcus aureus; 9, Micrococcus lysozyme; 10, Bacillus subtilis; 11, Escherichia coli; 12, Micrococcus luteus; 13, Edwardsiella tarda; 14, Salmonella paratyphosa; 15, Salmonella typhi; 16, Salmonella Schott; 17, Salmonella gallinarum; 18, Salmonella paratyphosa.

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Fig. 2.
The sensitivity of LAMP and PCR assays.
M, DL2000 DNA marker N: negative control. 1, 7.5 ng/μL; 2, 7.5×10-1 ng/μL; 3, 7.5×10-2 ng/μL; 4, 7.5×10-3 ng/μL; 5, 7.5×10-4 ng/μL; 6, 7.5×10-5 ng/μL; 7, 7.5×10-6 ng/μL; *, gDNA detection limit of LAMP or PCR assays. A, The sensitivity of LAMP assay; B, The sensitivity of PCR assay.
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Fig. 3.

the colony counts of 10-8 times diluted B. cereus. The average colony count of the three plates was 14 CFU/mL.

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Fig. 4.
results of enrichment time for detection of manually spiked contaminated samples. M, DL2000 DNA marker; N, negative control; 1, 15 min; 2, 30 min; 3, 45 min; 4, 60 min; *, indicated the shortest time for enrichment.
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Fig. 5.

The results of HNB used in LAMP assay. M, DL2000 DNA marker; 1, negative control; 2, HNB used in LAMP assay. A, indicated results of HNB in LAMP assay with naked eye; B, electrophoresis results of HNB in LAMP assay.

October 2023
Pakistan J. Zool., Vol. 55, Iss. 5, pp. 2001-2500

Pakistan Journal of Zoology

October

Pakistan J. Zool., Vol. 56, Iss. 5, pp. 2001-2500

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