In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

BACKGROUND: Extraction of cell-free DNA (cfDNA) is a key step for determining the quality of cfDNA-related molecular diagnostics. We evaluated the effect of sample containers and sample storage conditions on cfDNA extraction. METHODS: The cfDNA extraction using the MagMAX Cell-Free DNA Isolation Kit from five healthy controls and five lung cancer patients was evaluated according to the type of sample container and storage conditions: K2-EDTA container, <1, 6, 24, and 48 hr storage at 4℃ after immediate plasma separation; and Cell-Free DNA BCT container, <1, 3, 7, and 14 days stored at room temperature. Mutation analysis of EGFR exons 18–21 was performed. To assess the effect of a delay in centrifugation, EDTA whole blood samples from five healthy individuals were stored at 4℃ for 6, 12, and 24 hr before plasma separation. RESULTS: There was no significant difference in the amount and nucleic acid size of cfDNA in both controls and patients with cancer when EDTA plasma was stored at 4℃ up to 48 hr. The amount and size of cfDNA in the BCT container were not different up to 7 days; however, the 14-day sample showed an increase in cfDNA concentration due to genomic DNA contamination. EGFR mutations were detected on EDTA containers up to 48 hr and with BCT containers up to 14 days. When EDTA whole blood was stored at 4℃ and plasma separation was delayed, the cfDNA concentration increased from 24 hr. CONCLUSIONS: The cfDNA extraction was affected by the sample containers and storage conditions.
Biopsy ; Centrifugation ; DNA Contamination ; DNA ; Edetic Acid ; Exons ; Humans ; Lung Neoplasms ; Pathology, Molecular ; Plasma

Maca (Lepidium meyenii) is an herbaceous plant that grows in high plateaus and has been used as both food and folk medicine for centuries because of its benefits to human health. In the present study, ITS (internal transcribed spacer) sequences of forty-three maca samples, collected from different regions or vendors, were amplified and analyzed. The ITS sequences of nineteen potential adulterants of maca were also collected and analyzed. The results indicated that the ITS sequence of maca was consistent in all samples and unique when compared with its adulterants. Therefore, this DNA-barcoding approach based on the ITS sequence can be used for the molecular identification of maca and its adulterants.
DNA Barcoding, Taxonomic ; methods ; DNA, Intergenic ; analysis ; DNA, Plant ; analysis ; Drug Contamination ; prevention & control ; Humans ; Lepidium ; genetics ; Phytotherapy

The adulteration of herbal products is a threat to consumer safety. In the present study, we surveyed the species composition of commercial Radix Clerodendri Japonicum products using DNA barcoding as a supervisory method. A reference database for plant-material DNA-barcode was successfully constructed with 48 voucher samples from 12 Clerodendrum species. The database was used to identify 27 Radix Clerodendri Japonicum decoction piece samples purchased from drug stores and hospitals. The DNA sequencing results revealed that only 1 decoction piece (3.70%) was authentic C. japonicum, as recorded in the Dai Pharmacopeia, whereas the other samples were all adulterants, indicating a potential safety issue. The results indicate that decoction pieces that are available in the market have complex origins and that DNA barcoding is a suitable tool for regulation of Dai medicines.
Clerodendrum ; classification ; genetics ; DNA Barcoding, Taxonomic ; Drug Contamination ; Medicine, Chinese Traditional ; adverse effects ; Polymerase Chain Reaction

Adulteration in meat products is a widespread issue that could lead to serious threats to public health and religious violations. Technology that offers rapid, sensitive, accurate and reliable detection of meat species is the key to an effectual monitoring and control against meat adulteration. In recent years, high-throughput sequencing-based DNA metabarcoding technology has developed rapidly. With the characteristics of being high-throughput, highly precise and high-speed, this technology can simultaneously identify multiple species in complex samples, thus offering pronounced advantages in the surveillance of adulteration in meat and meat products. Starting with an introduction of the major developments in the high-throughput sequencing technology in the past two decades, this review provides an overview of the technical characteristics and research methods of DNA metabarcoding, summarizes the application of DNA metabarcoding technology in meat adulteration detection over the last few years, discusses the challenges of using DNA metabarcoding technology in the detection of meat adulteration, and provides future prospects on the development of this technology.
DNA ; Food Contamination/analysis* ; High-Throughput Nucleotide Sequencing/methods* ; Meat/analysis* ; Meat Products ; Technology

Ror fetal sex determination by the PCR method, oilgoprimers to Y- chromosome gene, DYZI, SRY, and AMGL were synthesized genomic DNA was extracted from male and female placenta for the control use. DYZI represented 154 bp single band to 0.001 pg/ml male genomic DNA but did not represent 154 bp band in female genomic DNA, SRY represented 341 bp bandto 1 pg/ml male genomic DNA in 2% agarose gel eleftrophoresis stained with ethidium bromide. DYZI was 1,000 fold sensitive than Sry and AMGL. DYZI and SRY could not identify the PDR failure from female but AMGL identified to 1,000-fold. During the dyal ampiification of female genomic DNA mixed with male genomic DNA, 0.00125 pg/nl, 1:400 part, male genomic DNA contamination represented male band but SRY amplification did not represent male band. It was suggested that SRY gene was deleted in two 46,XY felmle cases. For fetal sex determination, PCR with DYZL, SRY, and AMGL was performed in 10 cases. For fetal sex determination, PCR with DYZL, SRY, and AMGL with karyotyping in 10 cases of chorionic villi sex dietermination, PCR with DYZI, SRY, and AMGL was performed in 10 cases. For feral sex determination, PCR with DYZI, SRY, and AMGL with karyothping result, fetal sex determination, PCR with DYZI, SRY, and AMGL was performed in 10 Cases of choricinic villi and 15 cases of amnionic cells. By the comparison with karyotyping result, fetal sex determination was achieved successfully in all 23 samlies using PCR of SRY and AMGL but false result was detected in 3 cases(13%) using DYZI. Acording to our results, it was concluded that DYZL was 1,000-fold sensitive than SRY and AMGL but could not be used because of its false results, and AMGL and SRY must be used concomitantly for precise sex determination.
Amnion ; Chorionic Villi ; DNA ; DNA Contamination ; Ethidium ; Female ; Genes, sry ; Humans ; Karyotyping ; Male ; Placenta ; Polymerase Chain Reaction* ; Sepharose

OBJECTIVETo establish a high-quality method for detecting short tandem repeats(STR) using denaturing high performance liquid chromatography(DHPLC) in order to exclude maternal contamination and improve the accuracy of prenatal diagnosis.

METHODSTwo families were recruited. DNA was extracted from blood samples from the parents as well as amniotic fluid. Sixteen STR sites were amplified and analyzed based on the range of allele length reported by a STR database. Maternal DNA was mixed with DNA derived from amniotic fluid samples with the ratio 1:1, 1:4, 1:9, 1:19 and 1:99. vWA STR site was detected with DHPLC to confirm the sensitivity of detection.

RESULTSSixteen STR sites were analyzed by DHPLC, for which at least 10 were found to be different between the mothers and fetuses. The detection rate, with maternal contamination excluded, was 66.7%. And the sensitivity of detection was 1-10%.

CONCLUSIONMaternal contamination of amniotic fluid can be rapidly excluded with accuracy with DHPLC, which features a high sensitivity and good quality control, and can meet the European standards and provide a reliable quality control platform for prenatal diagnosis.

Chromatography, High Pressure Liquid ; methods ; DNA ; analysis ; chemistry ; genetics ; DNA Contamination ; Female ; Humans ; Microsatellite Repeats ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; methods

This research preliminarily discusses the relations of Dendrobium system growth through chloroplast gene rbcL, matK and the nuclear genome ITS2. The DNA barcoding universal sequence for authentication of the Dendrobium medical plants was slected and the possibility concerning utilizing the DNA barcoding to distinguish the D. huoshanenseand its adulterants was analyzed. Using the universal primer pair of ITS2, rbcL and matK, series of extended sequencing in the Dendrobium were conducted. Meanwhile, considering the different index about amplification and sequencing success rate of each sequence, the intraspecific and interspecific aberrance, the employment of BioEdit and MEGA 5.0 software were applied to establish the systematic tree of the NJ molecular and evaluate the diversified authentication capability of various sequences. The consequence demonstrates that the sequence of ITS2 is not only the largest one both in the intraspecific and interspecific aberrance of the Dendrobium but also has obvious barcoding gap. Considering the few overlap between the intraspecific and interspecific aberrance and the highest percentage regarding the formation of unilateral branch in diverse Dendrobium which have different ITS2 sequences, it can differentiate the species of Dendrobium. Furthermore, due to the inferior success rate of the rbcL and thematK and the lower reliability of NJ systematic tree, the percentage of the unilateral species which are generated by the systematic tree of rbcL and matK sequences is deficient. Therefore, the sequence of ITS2 can serves as DNA barcoding to distinguish the D. huoshanense, the D. moniliform and the D. officinale.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Dendrobium ; classification ; Drug Contamination ; Plant Preparations ; standards ; Plants, Medicinal ; classification ; Reproducibility of Results

OBJECTIVETo make the kit with witch to identify Penis et Testis Cervi with molecular taxonomy.

METHODThe mtDNA of sika and red deer from different areas was amplified by PCR and sequenced. Compared with the mtDNA of bovine and horse from witch the false medicines were made, characteristic segments of deer were found. We selected one as the species distinctive PCR primer of deer.

RESULTThe kit made up with this primer and related reagents could be used to discern Penis et Testis Cervi from the false medicine.

CONCLUSIONIt is a scientific, steady, accurate and convenient way to identify Penis et Testis Cervi with molecular taxonomy.

Animals ; Cattle ; genetics ; DNA ; genetics ; DNA Primers ; DNA, Mitochondrial ; genetics ; Deer ; classification ; genetics ; Drug Contamination ; Horses ; genetics ; Male ; Materia Medica ; chemistry ; Penis ; chemistry ; Testis ; chemistry

AIMTo study the difference of rDNA ITS sequences between Zanthexylum bungeanum populations and their adulterants in main habitants of China so as to provide molecular markers for identifying Zanthexylum bungeanum populations against adulterants.

METHODSrDNA ITS regions (including ITS-1, 5.8S and ITS-2) of 7 populations of Zanthexylum bungeanum which are separate located in Gansu, Shanxi, Sichuan, Hebei provinces, and 3 adulterants were sequenced by PCR products sequencing method or clone sequencing method.

RESULTSThe sequences of rDNA ITS region of Zanthexylum bungeanum were reported for the first time, and the sequences of ITS region ranged from 619 to 620 bp, and the length difference amoung Zanthexylum bungeanum and their adulterants is 4 bp. There are 15 variable sites, 12 informative sites and 3 authenticable sites among Zanthexylum bungeanum populations. The difference of rDNA ITS regions amoung Zanthexylum bungeanum and their adulterants is obvious, the number of variable sites is 71.

CONCLUSIONThe difference of rDNA ITS sequences can be used to authenticate accurately the populations of Zanthexylum bungeanum and their adulterants. These populations of Z. bungeanum which have close relationship always distribute in near geographic areas. The characteristics of rDNA ITS sequence can be used as good markers for authenticating Zanthexylum bungeanum populations form their adulterants.

Base Sequence ; China ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; Ecosystem ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; genetics ; Sequence Analysis, DNA ; Species Specificity ; Zanthoxylum ; classification ; genetics