1.PCR - based random amplified polymorphic DNA fingerprinting of vibrio cholerae 01 in Vietnam
Journal of Preventive Medicine 2003;13(5):27-30
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
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Random Amplified Polymorphic DNA Technique
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Vibrio cholerae
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DNA Fingerprinting
2.Application of random amplification polymorphic DNA in the genotyping of Neisseria gonorrhoeae.
Tie-jun ZHANG ; Yan-hua REN ; Ying-hua ZHANG ; Xiao-ming ZHOU ; Shun-zhang YU ; Qing-wu JIANG
Chinese Journal of Epidemiology 2004;25(9):779-782
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
3.Characterization of Sporothrix schenckii by random amplification of polymorphic DNA assay.
Xiaoming LIU ; Cuihong LIAN ; Liji JIN ; Lijia AN ; Guoling YANG ; Xiran LIN
Chinese Medical Journal 2003;116(2):239-242
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
4.Establishment of multiple regression model for virulence factors of Saccharomyces albicans by random amplified polymorphic DNA bands.
Qi LIU ; Youcong WU ; Youhua YUAN ; Li BAI ; Kun NIU
West China Journal of Stomatology 2011;29(6):643-645
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
5.Identification on Syngnathus and its adulterants with random amplified polymorphic DNA.
Yan WU ; Jia LIU ; Mengyue WANG ; Peijun JU ; Xiaobo LI
China Journal of Chinese Materia Medica 2009;34(14):1758-1760
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
6.Development and research advance of pharmacognosy field based on CNKI.
China Journal of Chinese Materia Medica 2018;43(4):689-695
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
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China
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DNA Barcoding, Taxonomic
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Data Mining
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Pharmacognosy
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Random Amplified Polymorphic DNA Technique
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Research
7.Genotypic Identification of Fusarium subglutinans, F. proliferatum and F. verticillioides Strains Isolated from Maize in Austria.
Youssuf A M H GHERBAWY ; Andereas ADLER ; Hansjorg PRILLINGER
Mycobiology 2002;30(3):139-145
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*
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DNA
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Fusarium*
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Gibberella
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Hand
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Random Amplified Polymorphic DNA Technique
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Zea mays*
8.RAPD analysis for genetic diversity of Fagopyrum cymosum.
Chunping ZHANG ; Ping HE ; Shijun HU ; Yifeng ZHANG ; Huacun JI ; Shan GAO
China Journal of Chinese Materia Medica 2009;34(6):660-663
OBJECTIVETo study the genetic diversity of Fagopyrum cymosum.
METHODThe genetic diversity of 87 individuals from 10 populations was analyzed by random amplified polymorphic DNA (RAPD).
RESULTTwelve primers were selected to produce highly reproducible RAPD bands. Among 85 amplified bands, seventy-five showed polymorphism, the percentage of polymorphic bands reached to 88.24%. Nei's gene diversity index (H) was 0. 3103, Shannon information index (I) was 0.5632, Gst was 0.1924. The genetic similarity coefficient and the genetic distance were 0.6720-0.9678 and 0.0328-0.3975, respectively.
CONCLUSIONF. cymosum shows high genetic diversity and the majority of genetic variation occurs in 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 F. cymosum.
Cluster Analysis ; Fagopyrum ; genetics ; Genetic Variation ; Phylogeny ; Random Amplified Polymorphic DNA Technique
9.RAPD analysis on genetic relationships of Erigeron breviscapus germplasm resources.
Sheng-Chao YANG ; Yong-Fang LI ; Zheng ZHAO ; Guo-Song WEN ; Yun LI ; You-Gui LIU ; Yong-Jian YANG ; Feng-Hui XIAO
China Journal of Chinese Materia Medica 2008;33(13):1532-1535
OBJECTIVETo investigate the genetic relationships of Erigeron breviscapus at the molecular biology level.
METHODThirty seven germplasm resources of E. breviscapus which collected from Yunnan, Sichuang and Guizhou province in 2005 were analyzed by Random amplified polymorphic DNA (RAPD) and cluster analysis based on NTSYS2.
RESULTA total of 10 primers were screened, and 107 bands were amplified, among which 94 (87.85%) bands were found to be polymorphic. Thirty seven germplasm resources of E. breviscapus were clustered into 3 groups at genetic distance 0.36, the I group include in 9 germplasm resources collected from Mile, Qiubei, Luxi, Gejiu, and Yanshan of south east of Yunnan province; the II group included 8 germplasm resources collected from Gucheng, and shangrila of north west of Yunnan province, and Mile and Qiubei of south east of Yunnan province; the III group included in 20 germplasm resources collected from the center of Yunnan province, north east of Yunnan province, Sichuan province, and Guizhou province.
CONCLUSIONThere were abundant genetic diversity in the germplasm resources of E. breviscapus, and the genetic relationships are closely related to geographical distance where they were collected.
Cluster Analysis ; Erigeron ; cytology ; genetics ; Genetic Variation ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique
10.Study on sequence characterized amplified region (SCAR) markers of Cornus officinalis.
Suiqing CHEN ; Xiaolei LU ; Lili WANG
China Journal of Chinese Materia Medica 2011;36(9):1145-1149
OBJECTIVETo establish sequence characterized amplified region markers of Cornus officinalis and provide a scientific basis for molecular identification of C. officinalis.
METHODThe random primer was screened through RAPD to obtain specific RAPD marker bands. The RAPD marker bands were separated, extracted, cloned and sequenced. Both ends of the sequence of RAPD marker bands were determined. A pair of specific primers was designed for conventional PCR reaction, and SCAR marker was acquired.
RESULTFour pairs of primers were designed based on the sequence of RAPD marker bands. The DNA of the seven varieties of C. officinalis was amplified by using YST38 and YST43 primer. The results showed that seven varieties of C. officinalis were able to produce a single PCR product. It was an effective way to identify C. officinalis. The varieties with cylindrical and long-pear shape fruits amplified by YST38 showed a specific band, which could be used as the evidence of variety identification. Seven varieties of C. oficinalis were amplified by using primer YST39. But the size of band of the variety with spindly shape fruit (35,0400 bp) was about 300 bp, which was shorter than those of the variety with the other shape fruits of C. officinalis (650-700 bp). The variety with the spindly shape fruit could be identified through this difference. The primer YST92 could produce a fragment from 600-700 bp in the varieties with cylindrical and long-pear shape fruits, a fragment from 200-300 bp in the varieties with oval and short-cylindrical shape fruits and had no fragment in the varieties with long cylindrical, elliptic and short-pear shape fruits, which could be used to select the different shapes of C. officinalis.
CONCLUSIONSCAR mark is established and can be used as the basis for breeding and distinguishing the verieties of C. officinalis.
Cornus ; classification ; genetics ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique