SNP genotyping of unpurified PCR products by sandwich-type affinity electrophoresis on a microchip with programmed autonomous solution filling

Lab on a Chip ◽  
2009 ◽  
Vol 9 (22) ◽  
pp. 3297 ◽  
Author(s):  
Akira Inoue ◽  
Aishan Han ◽  
Kimiko Makino ◽  
Kazuo Hosokawa ◽  
Mizuo Maeda
Keyword(s):  
Snp Genotyping ◽  
Affinity Electrophoresis ◽  
Pcr Products ◽  
Sandwich Type

A gel-free SNP genotyping method: bioluminometric assay coupled with modified primer extension reactions (BAMPER) directly from double-stranded PCR products

2004 ◽  
Vol 24 (2) ◽  
pp. 155-163 ◽  
Author(s):  
Guo-Hua Zhou ◽  
Hiromi Shirakura ◽  
Masao Kamahori ◽  
Kazunori Okano ◽  
Keiichi Nagai ◽  
...  
Keyword(s):  
Snp Genotyping ◽  
Primer Extension ◽  
Genotyping Method ◽  
Pcr Products

A Monoclonal Antibody-Based Enzyme Immunoassay for Fibrin Degradation Products in Plasma

1988 ◽  
Vol 59 (02) ◽  
pp. 310-315 ◽  
Author(s):  
P W Koppert ◽  
E Hoegee-de Nobel ◽  
W Nieuwenhuizen
Keyword(s):  
Enzyme Immunoassay ◽  
Degradation Products ◽  
Blood Clot ◽  
Tissue Type ◽  
Fibrin Degradation Products ◽  
Type Plasminogen Activator ◽  
Strong Positive Reaction ◽  
Sandwich Type ◽  
Capture Antibody

SummaryWe have developed a sandwich-type enzyme immunoassay (EIA) for the quantitation of fibrin degradation products (FbDP) in plasma with a time-to-result of only 45 minutes.* The assay is based on the combination of the specificities of two monoclonal antibodies (FDP-14 and DD-13), developed in our institute. FDP-14, the capture antibody, binds both fibrinogen degradation products (FbgDP) and FbDP, but does not react with the parent fibrin(ogen) molecules. It has its epitope in the E-domain of the fibrinogen molecule on the Bβ-chain between amino acids 54-118. Antibody DD-13 was raised using D-dimer as antigen and is used as a tagging antibody, conjugated with horse-radish peroxidase. A strong positive reaction is obtained with a whole blood clot lysate (lysis induced by tissue-type plasminogen activator) which is used as a standard. The EIA does virtually not detect FbgDP i. e. purified fragments X, Y, or FbgDP generated in vitro in plasma by streptokinase treatment. This indicates that the assay is specific for fibrin degradation products.We have successfully applied this assay to the plasma of patients with a variety of diseased states. In combination with the assay previously developed by us for FbgDP and for the total amount of FbgDP + FbDP (TDP) in plasma, we are now able to study the composition of TDP in patients plasma in terms of FbgDP and FbDP.

Abnormal Processing of the Glycoprotein IIb Transcript due to a Nonsense Mutation in Exon 17 Associated with Glanzmann’s Thrombasthenia

1995 ◽  
Vol 73 (05) ◽  
pp. 756-762 ◽  
Author(s):  
Yoshiaki Tomiyama ◽  
Hirokazu Kashiwagi ◽  
Satoru Kosugi ◽  
Masamichi Shiraga ◽  
Yoshio Kanayama ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Molecular Genetic ◽  
Genetic Defect ◽  
Nucleotide Sequence Analysis ◽  
Type I ◽  
Normal Amount ◽  
Immunoblot Assay ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Pcr Products

SummaryWe analyzed the molecular genetic defect responsible for type I Glanzmann’s thrombasthenia in a Japanese patient. In an immunoblot assay using polyclonal anti-GPIIb-IIIa antibodies, some GPIIIa (15% of normal amount) could be detected in the patient’s platelets, whereas GPIIb could not (<2% of normal amount). Nucleotide sequence analysis of platelet GPIIb mRNA-derived polymerase chain reaction (PCR) products revealed that patient’s GPIIb cDNA had a 75-bp deletion in the 3’ boundary of exon 17 resulting in an in-frame deletion of 25 amino acids. DNA analysis and family study revealed that the patient was a compound heterozygote of two GPIIb gene defects. One allele derived from her father was not expressed in platelets, and the other allele derived from her mother had a 9644C → T mutation which was located at the position -3 of the splice donor junction of exon 17 and resulted in a termination codon (TGA). Moreover, quantitative analysis demonstrated that the amount of the abnormal GPIIb transcript in the patient’s platelets was markedly reduced. Thus, the C → T mutation resulting in the abnormal splicing of GPIIb transcript and the reduction in its amount is responsible for Glanzmann’s thrombasthenia.

MOLECULAR STUDY OF HUMAN HEAD LICE, PEDICULUS HUMANUS CAPITIS COMPARISON WITH GOAT SUCKING LICE, LINOGNATHUS STENOPSIS (BURMEISTER) STRAINS

2020 ◽  
pp. 132-139
Keyword(s):  
Genetic Difference ◽  
Human Head ◽  
Molecular Study ◽  
Head Lice ◽  
Pediculus Humanus Capitis ◽  
School Students ◽  
Pediculus Humanus ◽  
Sucking Lice ◽  
Pcr Products ◽  
Amplicon Size

In this study, only (122) out of (915) primary school students were shown to be infected with head lice Pediculus. humanus capitis. The number and percentage of infected males were 46 (11.3%), while the number and percentage of infected females were 76 (14.9%). The results in our study also showed that the number and percentage of goats infected with goat sucking lice, Linognathus stenopsis was 70 (21.7%) of the total 322 animals, with the highest number and percentage among female goats 44 (62.9%) compared to the male goats 26 (37.1%). The study demonstrated that the rate of genetic difference between the studied samples was 89% and the similarity rate was 11%. Detection of OP-K01 gene pieces by PCR products showed that the amplicon size was 520 bp for P. humanus capitis isolated from humans, while the detection of OP-E20 and OP-M05 gene pieces with PCR product showed the lowest amplicon size 230 bp for Linognathus stenosis isolated from goats.

Spectrum of Defect States in Porous Organic Low-k Dielectric Films, Annealed in Argon and Nitrogen

2003 ◽  
Vol 766 ◽  
Author(s):  
V. Ligatchev ◽  
T.K.S. Wong ◽  
T.K. Goh ◽  
Rusli Suzhu Yu
Keyword(s):  
Deep Level ◽  
Dielectric Films ◽  
Silicon Substrates ◽  
Reverse Bias Voltage ◽  
Defect States ◽  
Single Side ◽  
Sandwich Type ◽  
Transient Spectroscopy ◽  
P Type ◽  
Spectrum Deconvolution

AbstractDefect spectrum N(E) of porous organic dielectric (POD) films is studied with capacitance deep-level-transient-spectroscopy (C-DLTS) in the energy range up to 0.7 eV below conduction band bottom Ec. The POD films were prepared by spin coating onto 200mm p-type (1 – 10 Δcm) single-side polished silicon substrates followed by baking at 325°C on a hot plate and curing at 425°C in furnace. The film thickness is in the 5000 – 6000 Å range. The ‘sandwich’ -type NiCr/POD/p-Si/NiCr test structures showed both rectifying DC current-voltage characteristics and linear 1/C2 vs. DC reverse bias voltage. These confirm the applicability of the C-DLTS technique for defect spectrum deconvolution and the n-type conductivity of the studied films. Isochronal annealing (30 min in argon or 60 min in nitrogen) has been performed over the temperature range 300°C - 650°C. The N(E) distribution is only slightly affected by annealing in argon. However, the distribution depends strongly on the annealing temperature in nitrogen ambient. A strong N(E) peak at Ec – E = 0.55 – 0.60 eV is detected in all samples annealed in argon but this peak is practically absent in samples annealed in nitrogen at Ta < 480°C. On the other hand, two new peaks at Ec – E = 0.12 and 0.20 eV appear in the N(E) spectrum of the samples annealed in nitrogen at Ta = 650°C. The different features of the defect spectrum are attributed to different interactions of argon and nitrogen with dangling carbon bonds on the intra-pore surfaces.

Linkage between H. pylori Infection and TNF-α polymorphism in The Pregnant Women

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Ghaidaa Raheem Lateef ◽  
Azhar Omaran Al-Thahab
Keyword(s):  
Pregnant Women ◽  
Serum Antibody ◽  
Rapid Test ◽  
Negative Group ◽  
Positive Group ◽  
Gynecology And Obstetrics ◽  
Tnf Α ◽  
Significant Difference ◽  
Pcr Products ◽  
H Pylori

A study was performed on 100 pregnant women in the outpatient department of gynecology and obstetrics of Maternity and Children Hospital in Al-Diwaniya City during the period between (March to September 2016). One hundred blood samples (50 for patients and 50 for control) were collected under the supervision of the treating gynecologist. The detection of Helicobacter. pylori was done by the use of the serum antibody Rapid test. The results showed that 50 (100%) were positive and 50 (100%) were negative for H. pylori in above method.All blood of patients and control samples were used for the extraction of genomic DNA,where the 107 bp PCR product size. Genotyping of the TNF-α-308 SNP (G/A)was performed by restriction fragment length polymorphism PCR (RFLP-PCR). PCR products were digested with restr NcoI iction enzyme. Individuals with the TNF-α-308(GG) homozygote produced digested DNA bands at 80,and 20 bp bp. A heterozygous genotype ofTNF-α-308 (GA)produced 107 bp,80 bp,and 20 bp bands. Individuals with the TNF-α-308 (AA) homozygote genotype had no amplicon digested and generated only one band of 107 bp. There was a significant difference in the frequency of the TNF-α-308(GG)genotype between H. pylori positive group and H. pylori negative group(72%,78% respectively). Also for GA genotype,there was a significant difference between H. pylori positive group and H. pylori negative group(24%,18% respectively). Concerning the frequency of the TNF-α-308 (AA)genotype between H. pylori positive group and H. pylori negative group,there was no significant difference between the two groups.

MICROSCOPIC AND MOLECULAR DIAGNOSIS OF Ascaridia spp. IN DOMESTIC PIGEONS(Columba livia domestica) IN BAGHDAD CITY,IRAQ

2020 ◽  
Vol 51 (4) ◽  
pp. 1220-1225
Author(s):  
Faraj & Al- Amery
Keyword(s):  
Molecular Diagnosis ◽  
Columba Livia ◽  
Ascaridia Galli ◽  
Molecular Assay ◽  
Infection Rates ◽  
Conventional Pcr ◽  
Significant Difference ◽  
Pcr Products ◽  
Males And Females ◽  
Domestic Pigeons

Ascaridiosis is a very important parasitic disease of birds, it is caused by Ascaridia. This study was conducted to identify the Ascaridia species by microscopic and molecular assay in Baghdad city. One hundred and sixty fecal samples were collected from domestic pigeons during the period from 1/1/ 2019 to 31/3/ 2019.  Results showed that the rate of infection for Ascaridia spp. 15.62% by microscopic examination.  Significant difference was observed in infection rates between males and females pigeons. Fifty samples randomly selected and subjected to molecular diagnosis of Ascaridia  spp.. Molecular examination results, the total infection rate showed 16%(8/50). The eight  positive PCR products were sequenced and deposited in Gene bank data base, phylogenic analysis demonstrated that 4 sequences belongs to Ascaridia galli ( MK918635.1, MK918636.1, MK918847.1, MK919081.1), while 2 (MK919199.1, MK919200.1) belong to  Ascaridia nymphii and 2 (MK919207.1, MK919264.1)  belong to Ascaridia numidae. It is the first study in Iraq to diagnosis of  Ascaridia nymphii and Ascaridia numidae  in domesticed pigeons by using conventional PCR.

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