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Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis

Esra Agel

Esra Agel

TUBITAK Marmara Research Center, Materials InstituteKocaeli, Turkey
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Agel, Esra
 y 
Hasan Sagcan

Hasan Sagcan

Istanbul Medipol University, Vocational School of Health Services, Department of Medical Laboratory TechniquesIstanbul, Turkey
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Sagcan, Hasan
  
21 oct 2020
Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis's Cover Image
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Categoría del artículo: Research Article
Publicado en línea: 21 oct 2020
Páginas: 230 - 236
DOI: https://doi.org/10.2478/ebtj-2020-0027
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© 2020 Esra Agel, Hasan Sagcan, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Results of different cryoprelyophilized mixture. 1) Freshly prepared mixture, 2) (0.5 % PEG 8000 + 2.0 % stachyose) Cryoprotectant mixture, 3) (0.5 % PEG 8000 + 1.0 % stachyose) Cryoprotectant mixture.
Results of different cryoprelyophilized mixture. 1) Freshly prepared mixture, 2) (0.5 % PEG 8000 + 2.0 % stachyose) Cryoprotectant mixture, 3) (0.5 % PEG 8000 + 1.0 % stachyose) Cryoprotectant mixture.

Figure 2

Results of lyophilized RT-PCR (a) and lyophilized LAMP (b) mixture. a: only 3. cryoprotectant mixture has positive result in RT-PCR assay. b: Results of LAMP assay 1) DNA ladder (100 bp, NEB), 2) 1. Cryoprotectant mixture, 3) 2. Cryoprotectant mixture, 4) 3. Cryoprotectant mixture 5) Freshly prepared mixture.
Results of lyophilized RT-PCR (a) and lyophilized LAMP (b) mixture. a: only 3. cryoprotectant mixture has positive result in RT-PCR assay. b: Results of LAMP assay 1) DNA ladder (100 bp, NEB), 2) 1. Cryoprotectant mixture, 3) 2. Cryoprotectant mixture, 4) 3. Cryoprotectant mixture 5) Freshly prepared mixture.

Figure 3

RT-PCR results of thermal stability of lyophilized mixtures
RT-PCR results of thermal stability of lyophilized mixtures

Figure 4

Detection limit results of lyophilized RT-PCR (a) and LAMP (b). a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control. b: a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control
Detection limit results of lyophilized RT-PCR (a) and LAMP (b). a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control. b: a: 1) 105 CFU/ml, 2) 104 CFU/ml, 3) 103 CFU/ml, 4) 102 CFU/ml, 5) 101 CFU/ml, 6) negative control

Figure 5

An image of lyophilized mixture in PCR tubes.
An image of lyophilized mixture in PCR tubes.

Cryoprotectant mixtures and amount of ingredients

The protocol of freeze dryer

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