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ARS Home » Pacific West Area » Riverside, California » National Clonal Germplasm Repository for Citrus » Research » Publications at this Location » Publication #299092

Title: A rapid field detection system for citrus huanglongbing associated ‘Candidatus Liberibacter asiaticus’ from the psyllid vector, diaphorina citri kuwayama and its implications in disease management

item Keremane, Manjunath
item RAMADUGU, CHANDRIKA - University Of California
item JENKINS, DANIEL - University Of Hawaii
item KUBOTA, RYO - University Of Hawaii
item SHIBATA, SCOTT - University Of Hawaii
item Hall, David
item ROOSE, MIKEAL - University Of California
item Lee, Richard
item RODRIGUEZ, ESTEBAN - University Of California

Submitted to: Crop Protection
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/28/2014
Publication Date: 11/28/2014
Citation: Keremane, M.L., Ramadugu, C., Jenkins, D., Kubota, R., Shibata, S., Hall, D.G., Roose, M., Lee, R.F., Rodriguez, E. 2014. A rapid detection system for citrus huanglongbing associated ‘Candidatus Liberibacter asiaticus’ from Diaphorina citri. Crop Protection. 68:41-48.

Interpretive Summary: We report the development of detection kit for the detection of the bacterium associated with huanglongbing (HLB). The sensitivity of detection is comparable to that of real time quantitative PCR. The kit may be operated in the field or in a field laboratory, does not require the use of expensive pipets, and does not require a highly skilled technician to operate. The unit can test up to eight samples simultaneously and is controlled by an Android smartphone or other device. The results, available in 30-40 minutes, are shown in real time. The availability of this affordable detection kit should be useful for farm managers, extension and regulatory personnel who can use timely diagnostic information to better manage HLB.

Technical Abstract: We report the development of an affordable detection kit for the detection of ‘Candidatus Liberibacter asiaticus’ (Las) from the psyllid vector, Diaphorina citri, which can provide real time test results in the field or field laboratory within 30-40 minutes without the need for expensive laboratory equipment or highly trained technical assistance. The kit uses the loop-mediated isothermal amplification (LAMP) technology which targets a portion of the Las genome other than the 16S rDNA region. The kit includes non-toxic buffer for extraction of DNA from the psyllids and the Smart-DART™ incubation unit. The Smart-DART™ unit can test eight different samples simultaneously and is operated by a Bluetooth connection with an Android device. The reaction results are shown in real time on the Android device and may be e-mailed for permanent storage and further analyses. Using psyllids from healthy and Las colonies, composite samples of 20 psyllids containing only one Las positive psyllid may be detected. This user friendly detection kit for Las in psyllids will be useful for growers in intra-orchard management, for extension and regulatory personnel, and for early detection of Las in areas where the D. citri is established but HLB is not widely present.