{"response":{"status":"ok","message_type":"publication"},"id":2609,"citation":"Louzada et al., 2022, HortScience","doi":"10.21273/hortsci16600-22","url":"https://api.seqco.de/v1/publications/2609.json","link_ext":"https://doi.org/10.21273/hortsci16600-22","title":"Optimization of vqPCR for Reliable Detection of Viable Candidatus Liberibacter asiaticus in Citrus","journal":"HortScience","journal_loc":"57 (6)","journal_date":"2022-06-01","pub_type":"journal-article","abstract":"\u003cjats:p\u003eCitrus Huanglongbing (HLB, also known as “citrus greening”), an important disease worldwide, is associated with three species of phloem-limited \u003cjats:italic\u003eCandidatus liberibacter\u003c/jats:italic\u003e, of which \u003cjats:italic\u003eCandidatus L. asiaticus\u003c/jats:italic\u003e (CLas) is the predominant one that has severely affected citrus production. TaqMan real-time polymerase chain reaction (PCR) (TM) has been the standard and very efficient method to diagnose several strains of Candidatus \u003cjats:italic\u003eLiberibacter\u003c/jats:italic\u003e in citrus; however, it detects total bacteria and is unable to differentiate dead from live \u003cjats:italic\u003eLiberibacter\u003c/jats:italic\u003e. The detection of only live bacteria is essential for testing methods of control for this important citrus disease. It is well known that ethidium monoazide and propidium monoazide (PMA) are compounds that supposedly enter only dead or membrane-damaged bacteria, intercalate the DNA strand, and make the DNA unavailable for amplification by PCR. These compounds are widely used when extracting the plant DNA to detect only live bacteria. In this research, we tested primers amplifying products from 79 to 1160 bp in TM and SYBR Green real-time PCR (SG) and PMA as DNA intercalating compound. Specifically, primers amplifying a 500-bp amplicon in SG provided the most reliable live-only detection, whereas those producing a smaller amplicon were unable to distinguish between live and dead. This is the first report of testing primers amplifying various amplicon sizes for the detection of only live CLas cells in citrus.\u003c/jats:p\u003e","long_citation_html":"Louzada et al. (2022). Optimization of vqPCR for Reliable Detection of Viable Candidatus Liberibacter asiaticus in Citrus. \n\u003ci\u003eHortScience\u003c/i\u003e. \u003ca href=\"https://doi.org/10.21273/hortsci16600-22\" target=\"_blank\"\u003eDOI:10.21273/hortsci16600-22\u003c/a\u003e\n","created_at":"2022-05-31T22:00:10.179Z","updated_at":"2025-11-06T13:54:26.735Z","authors":[{"id":10401,"given":"Eliezer","family":"Louzada","created_at":"2022-05-31T22:00:10.193Z","updated_at":"2022-05-31T22:00:10.193Z","url":"https://api.seqco.de/v1/authors/10401.json"},{"id":10402,"given":"Omar","family":"Vazquez","created_at":"2022-05-31T22:00:10.206Z","updated_at":"2022-05-31T22:00:10.206Z","url":"https://api.seqco.de/v1/authors/10402.json"},{"id":10403,"given":"Sandy","family":"Chavez","created_at":"2022-05-31T22:00:10.219Z","updated_at":"2022-05-31T22:00:10.219Z","url":"https://api.seqco.de/v1/authors/10403.json"},{"id":2358,"given":"Mamoudou","family":"Sétamou","created_at":"2019-04-15T18:47:17.508Z","updated_at":"2019-04-15T18:47:17.508Z","url":"https://api.seqco.de/v1/authors/2358.json"},{"id":2356,"given":"Madhurababu","family":"Kunta","created_at":"2019-04-15T18:47:17.494Z","updated_at":"2019-04-15T18:47:17.494Z","url":"https://api.seqco.de/v1/authors/2356.json"}],"names":[{"id":1,"name":"Candidatus Liberibacter asiaticus","url":"https://api.seqco.de/v1/names/1.json","uri":"https://seqco.de/i:1"},{"id":10973,"name":"Liberibacter","url":"https://api.seqco.de/v1/names/10973.json","uri":"https://seqco.de/i:10973"}],"subjects":[{"id":25,"name":"Horticulture","url":"https://api.seqco.de/v1/subjects/25.json"}]}