• 제목/요약/키워드: genotype identification

검색결과 141건 처리시간 0.021초

AP-PCR을 이용한 다제내성 Staphylococcus aureus의 유전형 분석 (Genotypic Analysis of Multi-drug Resistant Staphylococcus aureus by Arbitrarily Primed Polymerase Chain Reaction)

  • 신경현;홍승복;손승렬
    • 대한임상검사과학회지
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    • 제36권2호
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    • pp.89-97
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    • 2004
  • Many strains of Staphylococcus aureus were isolated from pus samples from primary, secondary, and tertiary medical institutions and were subjected to an antibiotic sensitivity test. Ciprofloxacin, clindamycin, erythromycin, gentamicin, oxacillin penicillin, tetracycline, trimethoprim/sulfamethoxazole, vancomycin and teicoplanin were used for the antibiotic sensitivity test. The strains showed hightest resistance to penicillin(91%), but all of strains tested were susceptible to vancomycin and teicoplanin. The isolated multi-drug(penicillin-tetracycline-ciprofloxacin-clindamycin-erythromycin- oxacillin-gentamicin) resistant S. aureus were analyzed genotypically using an AP-PCR(Arbitrarily Primed polymerase chain reaction) with an arbitrary 3 primers. Based on the result for genotype analysis, the genotypes identified by S1 primer did not coincide with those of S2 or E2 primers. Genotypes identified by S2 primer did not coincide with those of S1 or E2 primers. Also genotypes identified by the E2 primer did not coincide with those of S1 or S2 primers. Therefore, an analysis of AP-PCR test with multiple primers will provide more sensitive identification. A strain from a secondary medical institution and a strain from a tertiary medical institution which showed the same genotype for S1, S2, and E2 primers are required for further epidemiological study.

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A simple and rapid method for detection of single nucleotide variants using tailed primer and HRM analysis

  • Hyeonguk Baek;Inchul, Choi
    • 한국동물생명공학회지
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    • 제38권4호
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    • pp.209-214
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    • 2023
  • Background: Single nucleotide polymorphisms (SNPs) are widely used genetic markers with applications in human disease diagnostics, animal breeding, and evolutionary studies, but existing genotyping methods can be labor-intensive and costly. The aim of this study is to develop a simple and rapid method for identification of a single nucleotide change. Methods: A modified Polymerase Chain Reaction Amplification of Multiple Specific Alleles (PAMSA) and high resolution melt (HRM) analysis was performed to discriminate a bovine polymorphism in the NCAPG gene (rs109570900, 1326T > G). Results: The inclusion of tails in the primers enabled allele discrimination based on PCR product lengths, detected through agarose gel electrophoresis, successfully determining various genotypes, albeit with some time and labor intensity due to the use of relatively costly high-resolution agarose gels. Additionally, high-resolution melt (HRM) analysis with tailed primers effectively distinguished the GG genotype from the TT genotype in bovine muscle cell lines, offering a reliable way to distinguish SNP polymorphisms without the need for time-consuming AS-PCR. Conclusions: Our experiments demonstrated the importance of incorporating unique mismatched bases in the allele-specific primers to prevent cross-amplification by fragmented primers. This efficient and cost-effective method, as presented here, enables genotyping laboratories to analyze SNPs using standard real-time PCR.

Evaluation and Identification of Promising Bivoltine Hybrids of Silk-worm, Bombyx mori L., for Tropics

  • Rao, P.Sudhakara;Singh, Ravindra;Kalpana, G.V.;Naik, V.Nishitha;Basavaraja, H.K.;Swamy, G.N.Rama;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제3권1호
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    • pp.31-35
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    • 2001
  • Ten newly evolved bivoltine hybrids of silkworm (Bombyx mori L) were evaluated with control hybrid KA${\pm}$NB4D2 during three seasons of a year for their seasonal performance. Analysis of variance and other statistical methods were employed and the performance was observed in respect of 10 quantitative traits. The results showed significant genotype${\pm}$environment interaction with respect to four quantitative characters viz. fecundity, yield/10,000 larvae, filament length and raw silk (%). Environmental effects were significant for nine characters out of ten characters evaluated. A 105${\pm}$J2 and B${\pm}$NB4D2 were considered as highly adaptable hybrids to local conditions with high mean for maximum characters studied and found suitable to rear in all seasons.

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Main Gene Combinations and Genotype Identification of Hanwoo Quality with SNPHarvester

  • Bae, Jae-Young;Lee, Jea-Young
    • Communications for Statistical Applications and Methods
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    • 제19권6호
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    • pp.799-808
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    • 2012
  • It is known that human disease and the economic traits of livestock are significantly affected by a gene combination effect rather than a single gene effect. Existing methods to study this gene combination effect have disadvantages such as heavy computing, cost and time; therefore, to overcome those drawbacks, the SNPHarvester was developed to find the main gene combinations. In this paper, we looked for gene combinations using an adjusted linear regression model. This research finds that superior gene combinations which are related to the quality of the Korean beef cattle among sets of SNPs using SNPHarvester. We also identify the superior genotypes using a decision tree that can enhance the various qualities of Korean beef among selected a SNP combination.

An Improved PCR-RFLP Assay for Detection and Genotyping of Asymptomatic Giardia lamblia Infection in a Resource-Poor Setting

  • Hawash, Yoursry;Ghonaim, M.M.;Al-Shehri, S.S.
    • Parasites, Hosts and Diseases
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    • 제54권1호
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    • pp.1-8
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    • 2016
  • Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, $RIDA^{(R)}$ Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.

Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene

  • Pruthviraj, D.R.;Usha, A.P.;Venkatachalapathy, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권3호
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    • pp.315-320
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    • 2016
  • Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5'-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.

Identification of a Novel SNP Associated with Meat Quality in C/EBP${\alpha}$ Gene of Korean Cattle

  • Shin, S.C.;Kang, M.J.;Chung, E.R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권4호
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    • pp.466-470
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    • 2007
  • CCAAT/enhancer binding protein ${\alpha}$($C/EBP{\alpha}$) plays an important role in lipid deposition and adipocyte differentiation. In order to find genetic markers to improve the meat quality of Korean cattle, the bovine $C/EBP{\alpha}$ gene was chosen as a candidate gene to investigate its association with carcass and meat quality traits in Korean cattle. A single nucleotide polymorphism (SNP) was identified at position 271 (A/C substitution) of coding region in the $C/EBP{\alpha}$ gene. A PCR-RFLP procedure with restriction enzyme SmaI was developed for determining the marker genotypes. The frequencies of alleles C and A and were 0.374 and 0.626, respectively. The genotype frequencies for CC, AC and AA were 12.9, 49.0 and 38.1%, respectively, in Korean cattle population. The frequencies of genotype were in agreement with Hardy-Weinberg equilibrium. Association analysis indicated that the gene-specific SNP marker of $C/EBP{\alpha}$ showed a significant association with marbling score (p<0.05). The animals with AA genotype had higher marbling score than those with the AC or CC genotype. Although further studies are needed to validate our results, the $C/EBP{\alpha}$ gene could be useful as a genetic marker for carcass and meat quality traits in Korean cattle.

Meta-analysis of the Association Between GSTM1 and GSTT1 Gene Polymorphisms and Cervical Cancer

  • Zhang, Zhen-Yong;Jin, Xue-Ying;Wu, Rong;Wu, Li-Na;Xing, Rui;Yang, Shu-Juan;Xie, Yao
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권3호
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    • pp.815-819
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    • 2012
  • Aim: We conducted a meta-analysis to analyze the influence of GSTM1 and GSTT1 gene polymorphisms on cervical cancer risk, and explore gene-environment interactions. Methods: Identification of relevant studies was carried out through a search of Medline and the EMbase up to Oct. 2011. All case-control studies that investigated the association between GSTM1 and GSTT1 gene polymorphisms and risk of cervical cancer were included. The pooled odds ratio (OR) was used for analyses of results and the corresponding 95% confidence intervals (CI) were estimated. Results: A total of 21 case-control studies were included in the meta-analysis of GSTM1 (2,378 cases and 2,639 controls) and GSTT1 (1,229 cases and 1,223 controls) genotypes. The overall results showed that the GSTM1 null was related to an increased risk of cervical cancer (OR=1.50, 95% CI=1.21-1.85). Subgroup analysis were performed based on smoking and ethnicity. Our results showed that smokers with null GSTM1 genotype had a moderate increased risk of cervical cancer (OR=1.85, 95% CI=1.07-3.20). For the ethnicity stratification, moderate significantly increased risk of null GSTM1 genotype was found in Chinese (OR=2.12, 95% CI=1.43-3.15) and Indian populations (OR=2.07, 95% CI=1.49-2.88), but no increased risk was noted in others. Conclusion: This meta-analysis provided strong evidence that the GSTM1 genotype is associated with the development of cervical cancer, especially in smokers, and Chinese and Indian populations. However, no association was found for GSTT1 null genotype carriers.

Association of Polymorphisms in the Bovine Leptin Gene with Ultrasound Measurements for Improving in Korean Cattle

  • Kong, H.S.;Oh, J.D.;Lee, S.G.;Hong, Y.S.;Song, W.I.;Lee, S.J.;Kim, H.C.;Yoo, B.H.;Lee, H.K.;Jeon, G.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권12호
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    • pp.1691-1695
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    • 2006
  • The identification method that inflects real time ultrasound (RUT) and the potential application of marker assisted selection (MAS) for improvement of a cow population of Hanwoo (Korean Native cattle) was studied. The averages of RUT longissimus muscle area, RUT fat thickness, and RUT marbling score scanned at the 13th rib were 55.78 $cm^2$, 3.70 mm and 3.83 scores, respectively. We investigated the effects of the two SNPs (Kpn2 I and Msp I) in the leptin gene on carcass traits for Hanwoo cows by using ultrasound measurements. Genotype CC of the Kpn2 I had a significantly higher effect on back fat thickness (4.23 mm) and longissimus muscle area (57.57 $cm^2$) than genotype TT (3.14 mm, 53.93 $cm^2$, respectively, p<0.05). Genotype AA of the Msp I had a significantly higher effect only on marbling score (5.37) than genotype AB (3.57, p<0.05) and BB (3.37, p<0.05). Significant effects of SNPs in the leptin gene were found for the ultrasound measures of body composition in live cattle.

Microsatellite Marker를 이용한 한국재래돼지 집단의 품종특성 및 원산지 추적을 위한 개체식별체계 설정 (Characterization of a Korean Traditional Porcine Breed Using Microsatellite Markers and the Establishment of an Individual Identification System)

  • 김명직;이관호;오재돈;조규호;전기준;최봉환;이제현;홍윤숙;공홍식;이학교
    • 한국축산식품학회지
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    • 제27권2호
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    • pp.150-156
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    • 2007
  • 본 연구는 서로 다른 상염색체에 위치하고 있는 초위성체 유전 표지를 활용한 한국재래돼지 집단의 개체 식별시스템 설정을 위해 수행되었다. 공시재료는 4품종에서 총 446두가 사용되었으며 13종의 좌위에 대한 개체별 유전자형을 분석하였다. 이들 13종에서 출현된 이형접합도는 0.286-0.686였으며 marker 다형성 정보량은 0.399-0.796로 나타났다. 한국 재래돼지 집단에서 나타난 대립 유전자 발현 특성은 다른 대조 품종 집단과 매우 상이한 결과를 나타냈다. S0228좌위는 전체 8종의 대립유전자가 나타난 가운데 다른 3품종에서는 발현이 되지 않은 235 대립유전자가 한국 재래돼지 집단에서만 발현이 되었다. 5종의 초위성체 유전 표지를 활용할 경우 누적 개체 식별력은 99.999%를 나타냈으며 두 마리의 서로 다른 개체가 서로 같은 유전자형을 가질 짝확률은 $0.36{\times}10^{-9}$으로 추정되었다. 따라서 10종의 선정된 유전 표지는 한국재래돼지 집단에서 적정 신뢰도를 제공할 수 있는 개체 식별 시스템을 설정할 수 있을 것으로 생각된다.