Silicon Based System for Single-Nucleotide-Polymorphism Detection: Chip Fabrication and Thermal Characterization of Polymerase Chain Reaction Microchamber

The present study was carried out in the region of South Gujarat, India, to determine the prevalence and predisposing factors of canine parvovirus-2 (CPV-2) infection in acute gastroenteritis of pups. Further, haemagglutination (HA) test, enzyme linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) and rapid immunochromatography test were compared for diagnosis and prevalent CPV-2 types were ascertained. A total of 73 diarrhoea samples were collected and out of those 32, 33 and 35 were found positive via HA, ELISA and PCR, respectively. In rapid test, 26/52 samples were found positive. Among different age-groups, 11/24 and 13/21 animals were positive in pups aged 4–8 and 8–12 weeks, respectively. All but one (34/35) positive samples were from unvaccinated animals. Labrador was found to be the most susceptible breed (n = 13) to infection. Considering PCR as the best test, 47.94% (35/73) prevalence of CPV was recorded. Among PCR positive samples, 3 and 32 belonged to type CPV-2a and CPV-2b, respectively. Type CPV-2c was not detected among the examined samples. Sequencing analysis of 9/10 CPV-2b isolates revealed single nucleotide polymorphism (SNP) (A-G) at position 4106 (alanine to threonine) and suggested the occurrence of mutant, new CPV-2b in this area. As other major pathogen canine coronavirus was detected in 7/38 CPV negative samples. Conclusively, CPV-2 infection was detected in 47.97% cases of AGE of pups which warrants search for other pathogens in the diagnostic procedure. This work is among the few recent reports which depict the occurrence of a novel mutant (new CPV-2b) in India.

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Single nucleotide polymorphism detection method by temperature-gradient affinity chromatography using a single-stranded oligo-DNA coupled column

Journal of Chromatography A ◽

10.1016/j.chroma.2006.01.062 ◽

2006 ◽

Vol 1123 (2) ◽

pp. 182-188 ◽

Cited By ~ 2

Author(s):

Tomohisa Katsuda ◽

Ken Nishijima ◽

Mitsumasa Kamura ◽

Yasushi Nishiwada ◽

Shigeo Katoh

Keyword(s):

Single Nucleotide Polymorphism ◽

Temperature Gradient ◽

Affinity Chromatography ◽

Detection Method ◽

Nucleotide Polymorphism ◽

Single Nucleotide Polymorphism Detection ◽

Single Nucleotide ◽

Polymorphism Detection

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Identifikasi Keragaman Gen DGAT1 serta Asosiasinya terhadap Karakteristik Karkas dan Sifat Perlemakan Domba

Jurnal Ilmu dan Teknologi Peternakan Tropis ◽

10.33772/jitro.v6i2.7141 ◽

2019 ◽

Vol 6 (2) ◽

pp. 259

Author(s):

Asep Gunawan ◽

Ratna Sholatia Harahap ◽

Kasita Listyarini ◽

Cece Sumantri

Keyword(s):

Polymerase Chain Reaction ◽

Single Nucleotide Polymorphism ◽

Fragment Length Polymorphism ◽

Nucleotide Polymorphism ◽

Chain Reaction ◽

Single Nucleotide ◽

Dgat1 Gene ◽

Pcr Rflp ◽

Polymerase Chain ◽

Restriction Fragment Length

ABSTRAK Karakteristik karkas dan sifat perlemakan pada daging domba dikontrol oleh banyak gen salah satunya gen DGAT1 (Diacylglycerol Acyltransferasel 1). Penelitian ini bertujuan mengidentifikasi SNP (Single Nucleotide Polymorphism) gen DGAT1 pada titik mutasi g.8539 C>T dan asosiasinya terhadap karakteristik karkas dan sifat perlemakan pada domba Indonesia. Total sampel domba yang digunakan sebanyak 150 buah terdiri dari 35 sampel domba compass agrinak (DCA), 36 sampel domba barbados cross (DBC), 41 sampel domba komposit garut (DKG), 20 sampel domba ekor gemuk (DEG), dan 18 sampel domba ekor tipis (DET). Karakteristik karkas dan sifat perlemakan diukur dari domba jantan berumur 10-12 bulan. Identifikasi keragaman DGAT1|ALuI dianalisis dengan metode PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Hasil keragaman gen DGAT1 bersifat polimorfik dalam DET dan DEG, sedangkan DCA, DBC, dan DKG bersifat monomorfik. Dua genotipe disebut CC dan CT ditemukan dalam DET dan DEG. Titik mutasi gen DGAT1 berasosiasi (P<0.05) dengan karakteristik karkas, yaitu bobot dan panjang karkas. Selain itu, keragaman gen DGAT1 juga berasosiasi signifikan (P<0.05) dengan asam lemak jenuh, yaitu asam stearat (C18:0) dan asam arakidat (C20:0) dan asam lemak tak jenuh tunggal, yaitu asam oleat (C18:1n9c). Gen DGAT1 memiliki kontribusi dalam karakteristik karkas dan komposisi asam lemak pada domba.Kata Kunci: domba, gen DGAT1, karakteristik karkas, PCR-RFLP, sifat perlemakan ABSTRACT Characteristic of carcass and fatness traits of sheep is regulated by many genes such as DGAT1 (Diacylglycerol Acyltransferasel 1) gene. The research was aimed to investigate SNP (Single Nucleotide Polymorphism) of DGAT1 and its association with characteristic of carcass and fatness traits in Indonesian sheep. A total sample of sheeps used 150 rams of 10–12 months consisted 35 samples of compas agrinak sheep (CAS), 36 of barbados cross (BCS), 41 of garut composite (GCS), 20 of javanese fat tailed (JFT), and 18 of javanese thin tailed (JTT). Identification variant of DGAT1|ALuI were performed by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The results of polymorphism of DGAT1 were found in JTT and JFT. However, SNP of DGAT1 in CAS, BCS and GCS were monomorfic. Two genotype namely CC and CT were found in JTT and JFT populations. A SNP of the DGAT1 was associated (P<0.05) with characteristic of carcass, including weight and length of carcass. The variant of DGAT1 was associated too with saturated fatty acids (SFA) including stearic acid (C18:0) and arachidic acid (C20:0), and mono unsaturated fatty acid (MUFA) including oleic acid (C18:1n9c). The DGAT1 gene was contribute to characteristic carcass and fatty acid composition in sheep.Keywords: DGAT1 gene, characteristic carcass, fatness traits, PCR-RFLP, sheep

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