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. 2014 Nov 5;9(11):e112146.
doi: 10.1371/journal.pone.0112146. eCollection 2014.

Equipment-free incubation of recombinase polymerase amplification reactions using body heat

Affiliations

Equipment-free incubation of recombinase polymerase amplification reactions using body heat

Zachary Austin Crannell et al. PLoS One. .

Abstract

The development of isothermal amplification platforms for nucleic acid detection has the potential to increase access to molecular diagnostics in low resource settings; however, simple, low-cost methods for heating samples are required to perform reactions. In this study, we demonstrated that human body heat may be harnessed to incubate recombinase polymerase amplification (RPA) reactions for isothermal amplification of HIV-1 DNA. After measuring the temperature of mock reactions at 4 body locations, the axilla was chosen as the ideal site for comfortable, convenient incubation. Using commonly available materials, 3 methods for securing RPA reactions to the body were characterized. Finally, RPA reactions were incubated using body heat while control RPA reactions were incubated in a heat block. At room temperature, all reactions with 10 copies of HIV-1 DNA and 90% of reactions with 100 copies of HIV-1 DNA tested positive when incubated with body heat. In a cold room with an ambient temperature of 10 degrees Celsius, all reactions containing 10 copies or 100 copies of HIV-1 DNA tested positive when incubated with body heat. These results suggest that human body heat may provide an extremely low-cost solution for incubating RPA reactions in low resource settings.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Methods for securing tubes.
Mock RPA reactions were secured in the axilla using (A) a bandage, (B) an elastic sweatband, and (C) a strip of African chitenje fabric. An arrow is shown in each panel to indicate the approximate position of the tube, which is covered by material.
Figure 2
Figure 2. Temperature of mock reactions incubated at different body locations.
Each plot shows the temperature traces of mock RPA reactions incubated by 5 volunteers at 1 of 4 body location tested. Mock reactions were (A) held in the axilla, (B) taped to the abdomen, (C) placed in a rear trouser pocket, and (D) held in a closed fist.
Figure 3
Figure 3. Temperature of mock reactions secured to the body with different materials.
Each plot shows the temperature traces of mock RPA reactions incubated by 5 volunteers using 1 of 3 different materials. Materials tested included (A) a strip of cotton fabric, (B) a bandage, and (C) a sweatband.
Figure 4
Figure 4. Effect of ambient conditions on mock reaction temperature.
Each plot shows temperature traces of mock RPA reactions incubated under the arms of 5 volunteers in 1 of 4 environments with different ambient conditions. Mock reactions were incubated (A) in a cold room at 4 degrees Celsius, (B) in a cold room at 10 degrees Celsius, (C) at room temperature, and (D) in the Houston summer sun.

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