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[Authentication of Lonicera japonica using bidirectional PCR amplification of specific alleles]
Chao Jiang1, Ya-Hua Zhang, Min Chen
1Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Objective:
To identify SNP in flos Lonicerae, and authenticate Lonicera japonica from its adulterants and the mixture by using bidirectional PCR amplification of specific alleles (Bi-PASA).
Method:
SNP of L. japonica and its adulterants was identified by using ClustulW to align trnL-trnF sequences of the Lonicera genus from GenBank database. Bi-PASA primer was designed and the PCR reaction systems including annealing temperature optimized. Optimized result was performed in 84 samples to authenticate L. japonica, its adulterants and the mixture.
Result:
When the annealing temperature was 61 degrees C, DNA from L. japonica would be amplified 468 bp whereas PCR products from all of the 9 adulterants were 324 bp. The established method also can detect 5% of intentional adulteration DNA into L. japonica.
Conclusion:
The Bi-SPASA could authenticate L. japonica from its adulterants and the mixture.
