Genetic Identification Of Edible Bird’s Nest In Thai Land And Quick Detection Technique Method

ObjectiveEdible bird’s nest species of Aerodramus genus in the Apodidae family,Swiftlet many of swallows of Aerodramus genus.In Thailand have three species of edible bird’s nest.Edible-nest swiftlet(Aerodramus fuciphagus)and White-rumped swiftlet(Aerodramus fuciphagus germani).Black-nest swiftlet(Aerodramus maxima).Swiftlet habitat in natural caves or similar caves and can be found in central,eastern and southern in Thailand.Bird’s nest is mostly produced in Indonesia,Thailand,Vietnam,Malaysia and Singapore.The bird’s nest production in Thailand accounts for about 5%of the world’s production,mainly distributed in central,eastern and southern Thailand.According to the different living environment of Swiftlet,bird’s nest divided into cave bird’s nest and house bird’s nest.At present,at present,there are more researches on bird’s nest origin in Indonesia and Malaysia,and less research on bird’s nest origin in Thailand.The use of modern molecular biology techniques to identify the original bird’s nest can be used as an effective supplement to traditional experience identification.I am foreign students from Thailand.I collected 30 bird’s nest specimens in different regions of Thailand during the summer vacation,26 of which were house swallows and 4 were cave swallows.This study uses modern molecular biology techniques to identify the origin of bird’s nest samples in different regions of Thailand,and provides basic data for the distribution of bird’s nest resources in Thailand.Methods1.Comparsion of the four methods for DNA extraction of Edible bird’s nest;CTAB method,SDS method,oral pharyngeal swab gene DNA extraction kit,Bird’s Nest Nucleic Acid Extraction Kit.The extraction method is simple,fast,and time-saving.2.Edible bird’s nest samples collect total 30 kinds from different parts of Thailand:Eastern,Western,and Southern.There are 26 samples of house swallows and 4 samples of cave swallows.Cytb gene sequences were used to amplify the length by PCR and the sample sequences analyzed by gel-electrophoresis.Analysis of the BLAST results on the NCBI website,downloading the database of the Cytb gene sequence of Aerodramus in GenBank,and using MEGA-X software to perform an NJ construction tree comparison on the sequences of all Edible bird’s nest samples to analyze the relationship.3.Use the ND2 gene sequence to identify the bird’s nest bio-based prototyping for 30 samples of bird’s nest in Thailand.Download the data of the ND2 gene sequence of Aerodramus fuciphagus on NCBI website to design primers and perform PCR amplification and sequencing.Use BLAST search for similarities.4.The ARMS-PCR method was established to identify origin of bird’s nest and the rapid identification technology of bird’s nest in Thailand was discussed.The first primer is used to identify Aerodramus fuciphagus in the bird’s nest,and the second primer is used to identify the Aerodramus maximus in the bird’s nest.Results1.Four methods for DNA extraction of Edible bird’s nest were compared by investigating sample of nucleic acid purity,electrophoresis band and operating time.results sample nucleic acid purity:Bird’s Nest Nucleic Acid Extraction Kit>CTAB method>SDS method>oral pharyngeal swab gene DNA extraction kit;2.Cytb gene sequence identification was based on the original Thailand bird’s nest bio-results.The results of similarity of Thai bird’s nest samples in different regions in BLAST search.results.All the Thai bird’s nest samples have the highest sequence similarity to Aerodramus fuciphagus.Results in NJ Tree showed that 26 samples of house nest are distributed in central,western,eastern and southern parts of Thailand was Aerodramus fuciphagus.4 sample of cave bird’s nest in southern Thailand was Aerodramus Maximus.3.ND2 gene sequence identification was based on the Thai bird’s nest bio-based original results.The Thai bird’s nest samples in different regions were searched for similarity in BLAST results.All the Thailand bird’s nest samples had the highest sequence similarity to Aerodramus fuciphagus and Aerodramus Maximus.The results of building trees in NJ showed that 26 samples samples of house nest are distributed in central,western,eastern and southern parts of Thailand was Aerodramus fuciphagus,4 samples of cave bird’s nest in southern Thailand was Aerodramus Maximus.4.The ARMS-PCR method,identification two kinds of bird’s nest.The first primers used to identify Aerodramus fuciphagus and Aerodramus maximmus.Identify Aerodramus fuciphagus,PCR amplification via primers Af/2-693bp-F/R、Af/2-A-249bp-F、Af/2-G-498bp-R,on electrophoresis agarose gel had band lengths of 693bp,498bp,249bp.Identify Aerodramus maximmus,PCR amplification via primers Am-ND2-463bp-F/R、Am-ND2-A-317bp-F、Am-ND2-G-201bp-R,had band lengths of 463bp,317bp and 201bp in the electrophoresis gel.Conclusion1.Four methods for DNA extraction of Edible bird’s nest(CTAB method,SDS method,oral pharyngeal swab gene DNA extraction kit,Bird’ s Nest Nucleic Acid Extraction Kit method),Bird’s Nest Nucleic Acid Extraction Kit is the best extraction method.2.Based on the Cytb gene,the NJ tree was constructed after sequencing.The identification results showed that the bird’s nest in Thailand is dominated by two types of birds,Aerodramus fuciphagus and Aerodramus Maximus.3.Based on the ND2 gene,the NJ tree was constructed after sequencing.The identification results showed that the bird’s nest in Thailand is dominated by two types of birds,Aerodramus fuciphagus and Aerodramus Maximus.4.Using ARMS-PCR detection method to design specific primers for bird’s nest primitive identification,which can be quickly identified.