Nested PCR AP Group - 24 tests / Nested PCR AP Group - 96 tests

Detection: Candidatus Phytoplasma mali, Candidatus Phytoplasma prunorum and Candidatus Phytoplasma pyri without distinction.

Matrices: Symptomatic leaf mid-vein tissue and/or vascular tissue (phloem) from bark or roots. Analysis on asymptomatic plants is possible but not recommended.

Other information: Kit developed and optimized by Qualiplante according to a method published in the Appendix 4 of PM7/62 (3) ‘Candidatus Phytoplasma mali’, ‘Ca. P. pyri’ and ‘Ca. P. prunorum’, European and Mediterranean Plant Protection Organization Bulletin (2020) 50 (1), 69–85. Permits to detect the following phytoplasmas Candidatus Phytoplasma mali responsible of Apple Proliferation (AP) on apple tree, Candidatus Phytoplasma prunorum responsible of European Fruit Stone Yellow (EFSY) on apricot tree and Candidatus Phytoplasma pyri responsible of Pear Decline (PD) on pear tree. Only an enzymatic digestion permits to distinguish the different phytoplamas of Apple Proliferation group (reagents not included with the kit).

The method is based on a first amplification of P1/P7 primers (Deng & Hiruki, 1991; Schneider et al., 1995) that permits to confirm the presence of a phytoplasma amplifying the whole length of 16S and intergenic 16S-23S and a small part of 23S rRNA gene. FO1/rO1 primers (Lorenz et al. 1995) are used for second PCR and are specific for 16SrX group phytoplasmas.

Validation data of the method are available from a test performance study realized in 2011 (Euphresco FruitPhytoInterlab project), where 20 laboratories analysed a total of 30 samples. The performance characteristics obtained are:

  • Analytical sensitivity for ‘ P. mali’: 101 – 103
  • Analytical sensitivity for ‘ P. pyri’: 101
  • Diagnostic sensitivity: 99.3%
  • Diagnostic specificity: 97.7%
  • Repeatability: no data available
  • Reproducibility: Kappa coefficient 0.94

Included reagents: Direct Master Mix, Nested Master Mix, Negative Control, Positive Control.

 

Targeted pathogen:


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