The random amplified polymorphic DNA (RAPD) technique was used to distinguish 60 strains of Vibrio cholerae 01 E1 Tor isolated from patients in 1991-2002 period. There were 6 strains isolated from HoChiMinh city, 43 strains from Nha Trang, 8 strains from Hanoi and 3 strains from Tay Nguyen Highland. In RAPD technique, DNA oligomers with 10 nucleotids was selected randomly using to synthese prime DNA from the sites of properly paired aminoacids. Strain specific amplified DNA was created. AP42 and AP46 were used to identify the gene of various strains in diverse areas. Results of RAPD showed that these strains of bacteria had a same origin. RAPD technique can be used to supervise the transmission of bacteria in human
DNA ; Random Amplified Polymorphic DNA Technique ; Vibrio cholerae ; DNA Fingerprinting

OBJECTIVETo set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection.

METHODSFour different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains. Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared.

RESULTSCetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar.

CONCLUSIONBased on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.

DNA Fingerprinting ; DNA, Bacterial ; analysis ; Genotype ; Humans ; Neisseria gonorrhoeae ; classification ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo study the identification method of Syngnathus and its adulterants.

METHODRandom amplified polymorphic DNA (RAPD) was used to construct a dendrogram by UPGMA method based on Nei & Li's coefficient and a genetic affinity pattern for Syngnathus acus, Solenognathus hardwickii, Syngnathoides biaculeatus, Trachyrhamphus serratus, Halicampus koilomatodon, Microphis boaja.

RESULTFour primers, LJ04, LJ09, LJ16 and LJ19, from 18 random primers were used in the dendrogram which can differentiate Syngnathus in genus level and showed a great consistence with the appearance identification. The genetic affinity pattern based on primers LJ09 and LJ19 could be used to identify Syngnathus from its adulterants.

CONCLUSIONRAPD is suitable to identify Syngnathus and its adulterants.

Animals ; China ; DNA Primers ; genetics ; Phylogeny ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; Smegmamorpha ; classification ; genetics

OBJECTIVETo research the relationship between the virulence factors of Saccharomyces albicans (S. albicans) and the random amplified polymorphic DNA (RAPD) bands of them, and establish the regression model by multiple regression analysis.

METHODSExtracellular phospholipase, secreted proteinase, ability to generate germ tubes and adhere to oral mucosal cells of 92 strains of S. albicans were measured in vitro; RAPD-polymerase chain reaction (RAPD-PCR) was used to get their bands. Multiple regression for virulence factors of S. albicans and RAPD-PCR bands was established.

RESULTSThe extracellular phospholipase activity was associated with 4 RAPD bands: 350, 450, 650 and 1 300 bp (P < 0.05); secreted proteinase activity of S. albicans was associated with 2 bands: 350 and 1 200 bp (P < 0.05); the ability of germ tube produce was associated with 2 bands: 400 and 550 bp (P < 0.05).

CONCLUSIONSome RAPD bands will reflect the virulence factors of S. albicans indirectly. These bands would contain some important messages for regulation of S. albicans virulence factors.

DNA ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique ; Saccharomyces ; Virulence Factors

OBJECTIVESTo investigate the DNA polymorphism of Sporothrix schenckii (S. schenckii) and to find the relationship between DNA patterns and geographic areas and clinical manifestations.

METHODThe total DNA was extracted with hexadecyltrimethyl-ammonium bromide. Random Amplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S. schenckii collected from different areas and isolated from different clinical types.

RESULTSOf seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5'-ACCCGACCTG-3', 5'-GAGAGCCAAC-3', 5'-GGTGAC~GCAG-3' respectively. The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability. Different isolates showed a common conserved DNA band with the same primer. Different clinical types showed different genotypes.

CONCLUSIONRAPD analysis is useful in DNA typing of S. schenckii, the DNA band type of which is related to geographic origin and Clinical manifestation.

DNA, Fungal ; analysis ; Humans ; Random Amplified Polymorphic DNA Technique ; Sporothrix ; genetics

Based on the literature data in CNKI, data mining and analysis technologies were used in this paper to describe the scientific research and development direction of Pharmacognosy in the last decade from the perspective of bibliometrics. The analysis of measured data revealed the core research institutions, excellent research teams, leading scholars, major research aspects and research progress in the field. Results showed that most of the scholars in the field were from colleges and institutions, accounting for 74.6% of the total research findings and forming a group of core scholars. In terms of frequency and timeliness of citation, pharmacognosy is a discipline in sustained growth and development since it mainly cites the literature in the other disciplines, absorbs and utilizes knowledge of the other disciplines. Over the last few years, molecular identification and genetic diversity have become the research hotspots in pharmacognosy, and the techniques and methods such as ISSR, RAPD, DNA barcoding and DNA molecular marker have been widely used.
Bibliometrics ; China ; DNA Barcoding, Taxonomic ; Data Mining ; Pharmacognosy ; Random Amplified Polymorphic DNA Technique ; Research

Gibberella fujikuroi is species complex. This species complex includes Fusarium tabacinum, F. moniliforme (= F. verticillioides), F. nygamai, F. proliferatum as well as F. subglutinans. Our objective was to develop a technique to differentiate between isolates of F. subglutinans, F. proliferatum and F. verticillioides. Thirty-two strains of F. subglutinans, six strains from F. verticillioides and five strains of F. Proliferatum isolated from maize in Austria were studied using random amplified polymorphic DNA (RAPD). F. subglutinans strains clustered very closely, with similarity ranging from 87~100%. On the other hand, all the amplification patterns of F. verticillioides were identical, as well as in the case of F. proliferatum. Our results indicated that these Fusaria species are distinct species and hence RAPD markers can be quick and reliable for differentiating them.
Austria* ; DNA ; Fusarium* ; Gibberella ; Hand ; Random Amplified Polymorphic DNA Technique ; Zea mays*

OBJECTIVETo establish an effective way for rapid identification of Monascus strains based on DNA molecular marker.

METHODA random amplified polymorphic DNA (RAPD) marker named F421 in genomic DNA of Monascus F strain was observed during a comparison of DNA fingerprints derived from 10 cultivated strains of Monascus. F421 was cloned and sequenced. Comparing the sequence of F421 (GenBank accession number EF063107) with other relative sequences in the GenBank databases, no distinct comparability was found. A pair of sequence characterized amplified region (SCAR) primers were designed based on the sequence of the cloned fragment and tested for the specific detection of Monascus F.

RESULTThe results of polymerase chain reaction showed that only a 421bp segment of Monascus F strain was amplified compared with other 9 cultivated strains of Monascus. And the acquired SCAR marker of strain F could be used as a specific DNA fingerprint to identify Monascus strain F within one day.

CONCLUSIONSCAR molecular marker technology is an effective new way to identify Monascus strains more rapidly. And also is an assistant tool to identify Monascus strains more accurately when disagreements come out using traditional classification. It could be applied widely to the protection of germ plasm resources, classification and identification distinguishing false strains of pharmaceutical fungi.

Molecular Sequence Data ; Monascus ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo analyze the genetic diversity of wild Rehmannia glutinosa and evaluate and compare random amplified polymorphic DNA (RAPD) and inter sample sequence repeat (ISSR) for analysis of R. glutinosa accessions.

METHODTwo molecular markers, RAPD and ISSR were used for analyzing 55 wild R. glutinosa accessions.

RESULTAverage 16.00 and 19.08 bands were amplified by RAPD primers and ISSR primers respectively, and the percentage of polymorphic bands were 89.58% and 94.32% respectively; Fifty-five R. glutinosa accessions categorized into 7 clusters were identified by unweighted pair-group method, arithmetic average (UPGMA) method.

CONCLUSIONA high level of genetic diversity of wild Rehmannia glutinosa was displayed at DNA level, and genetic diversity coefficient of R. glutinosa from different production areas was 0.63-0.98, and ISSR marker can detect higher genetic diversity of R. glutinosa germplasms than RAPD marker.

Genetic Variation ; genetics ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; methods ; Rehmannia ; classification ; genetics

OBJECTIVETo discuss the genetic diversity of Coptis omeiensis.

METHODThe genetic diversity of 110 individuals from 10 populations was analyzed by RAPD.

RESULT14 primers were selected to produce highly reproducible RAPD bands. Among 132 amplified bands, 98 showed polymorphism, the percentage of polymorphic bands reached to 74.24%. Nei's gene diversity index (H) was 0.2863, Shannon's information index (I) was 0.3624, G(st) was 0.2305. The genetic distance coefficient and the similarity were 0.1931-0.5245 and 0.5016-0.8843, respectively.

CONCLUSIONThere exists a held high genetic diversity in C. omeiensis and the majority of genetic variation occurs in the populations. By cluster analysis, the geographical distribution is very obvious. The RAPD marker can be used for the analysis of the genetic diversity and genetic variation of C. omeiensis.

China ; Coptis ; classification ; genetics ; Genetic Variation ; Phylogeny ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique