Authentication of Lonicera japonica using bidirectional PCR amplification of specific alleles / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>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).</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>The Bi-SPASA could authenticate L. japonica from its adulterants and the mixture.</p>