Evaluation and usefulness of reverse dot blot DNA-PolyMarker typing in forensic case work

1996 ◽  
Vol 81 (1) ◽  
pp. 61-72 ◽  
Author(s):  
N. Dimo-Simonin ◽  
C. Brandt-Casadevall
Keyword(s):  
Dot Blot ◽  
Forensic Case ◽  
Case Work ◽  
Reverse Dot Blot

APPLICATION OF REAL-TIME PRC TECHNIQUE AND REVERSE DOT-BLOT FOR DETECTION THE HIGH RISK TYPES OF HUMAN PAPILLOMAVIRUS CAUSING CERVICAL CANCER

2017 ◽  
pp. 99-103
Author(s):  
Van Bao Thang Phan ◽  
Hoang Bach Nguyen ◽  
Van Thanh Nguyen ◽  
Thi Nhu Hoa Tran ◽  
Viet Quynh Tram Ngo
Keyword(s):  
Cervical Cancer ◽  
High Risk ◽  
Real Time ◽  
Real Time Pcr ◽  
Dot Blot ◽  
The Real ◽  
Dot Blot Assay ◽  
Hpv Types ◽  
High Risk Hpv ◽  
Reverse Dot Blot

Introduction: Infection with HPV is the main cause of cervical cancer. Determining HPV infection and the types of HPV plays an important role in diagnosis, treatment and prognosis of cervicitis/cervical cancer. Aims: Determining proportion of high-risk HPV types and the occurrence of coinfection with multiple HPV types. Methods: 177 women with cervicitis or abnormal Pap smear result were enrolled in the study. Performing the real-time PCR for detecting HPV and the reverse DOT-BLOT assay for determining type of HPV in cases of positive PCR. Results: 7 types of high-risk HPV was dectected, the majority of these types were HPV type 18 (74.6%) and HPV type 16 (37.6%); the proportion of infection with only one type of HPV was 30.4% and coinfection with multiple HPV types was higher (69.6%), the coinfected cases with 2 and 3 types were dominated (32.2% and 20.3%, respectively) and the coinfected cases with 4 and 5 types were rare. Conclusion: Use of the real-time PCR and reverse DOT-BLOT assay can determine the high-risk HPV types and the occurrence of coinfection with multiple HPV types. Key words: HPV type, Reverse DOT-BLOT, real-time PCR,PCR, cervical cancer

Simplified reverse dot blot analyses for detecting ras oncogene mutations

1997 ◽  
Vol 2 (3) ◽  
pp. 169-176 ◽  
Author(s):  
M ALBITAR ◽  
W WU ◽  
E FELTZ ◽  
G JIN ◽  
C HIRSCHGINSBERG ◽  
...  
Keyword(s):  
Ras Oncogene ◽  
Dot Blot ◽  
Reverse Dot Blot

Hybridoma Screening by Antigen Capture: Reverse Dot Blot

2022 ◽  
Vol 2022 (1) ◽  
pp. pdb.prot103135
Author(s):  
Edward A. Greenfield
Keyword(s):  
Tissue Culture ◽  
Monoclonal Antibodies ◽  
Culture Supernatant ◽  
Membrane Surface ◽  
Nitrocellulose Membrane ◽  
Dot Blot ◽  
Tissue Culture Supernatant ◽  
Antigen Capture ◽  
Mouse Immunoglobulin ◽  
Reverse Dot Blot

A dot blot is widely used to determine the productivity of a given hybridoma. This assay can also be used to screen a fusion or subclone plate for productive hybridoma clones. First, a nitrocellulose membrane is coated with an affinity-purified goat or rabbit anti-mouse immunoglobulin and then incubated with hybridoma tissue culture supernatant. Monoclonal antibodies in the supernatant are then “captured” on the coated nitrocellulose membrane surface and detected by screening with horseradish peroxidase (HRP).

scholarly journals Alpha and beta-Thalassemia mutations in Hubei area of China

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Yaowu Zhu ◽  
Na Shen ◽  
Xiong Wang ◽  
Juan Xiao ◽  
Yanjun Lu
Keyword(s):  
Mean Corpuscular Volume ◽  
Southern China ◽  
Hubei Province ◽  
Beta Thalassemia ◽  
Corpuscular Volume ◽  
Dot Blot ◽  
Inherited Disorders ◽  
Great Heterogeneity ◽  
Reverse Dot Blot

Abstract Background Thalassemia is a group of inherited hemoglobic disorders resulting from defects in the synthesis of one or more of the hemoglobin chains, which is one of the most prevalent inherited disorders in southern China. Only few studies reported the molecular characterization of α- and β-Thalassemia in Hubei Province in the central of China. Methods A total of 4889 clinically suspected cases of thalassemia were analyzed by Gap-PCR, PCR-based reverse dot blot (RDB). Results 1706 (33.8%) subjects harbored thalassemia mutations, including 539 (11.0%) subjects with α-thalassemia, 1140 (23.3%) subjects with β-thalassemia mutations, and 25 (0.51%) subjects with both α- and β-thalassemia mutations. Seven genotypes of α-thalassemia mutations and 29 genotypes of β-thalassemia mutations were characterized. --SEA/αα (66.05%), −α3.7/αα (24.12%), and -α4.2/αα (3.71%) accounted for 93.88% of the α-thalassemia mutations. βIVS-II-654/βN, βCD41–42/βN, βCD17/βN, βCD27–28/βN, βCD71–72/βN, β − 28/βN, β − 29/βN, βCD43/βN, βE/βN, accounting for 96.40% of all β-thalassemia genotypes. Furthermore, mean corpuscular volume (MCV) and mean corpuscular Hb (MCH) were sensitive markers for both β-thalassemia and α-thalassemia with --SEA/αα, but not -α3.7/αα and -α4.2/αα. Conclusions: Our data indicated great heterogeneity and extensive spectrum of thalassemias in Hubei province of China.

Nearly 80% of cystic fibrosis heterozygotes and 64% of couples at risk may be detected through a unique screening of four mutations by ASO reverse dot blot

Genomics ◽  
1991 ◽  
Vol 11 (4) ◽  
pp. 1149-1151 ◽  
Author(s):  
J.L. Serre ◽  
A. Taillandier ◽  
E. Mornet ◽  
B. Simon-Bouy ◽  
J. Boué ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
At Risk ◽  
Dot Blot ◽  
Reverse Dot Blot

Prenatal diagnosis of β-thalassaemia by reverse dot-blot hybridization

1999 ◽  
Vol 19 (5) ◽  
pp. 428-435 ◽  
Author(s):  
Pranee Winichagoon ◽  
Vannarat Saechan ◽  
Roongrat Sripanich ◽  
Chamnong Nopparatana ◽  
Sujin Kanokpongsakdi ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Reverse Dot Blot Hybridization ◽  
Reverse Dot Blot

scholarly journals Reverse dot-blot detection of the African-American beta-thalassemia mutations

Blood ◽  
1995 ◽  
Vol 86 (4) ◽  
pp. 1580-1585 ◽  
Author(s):  
P Sutcharitchan ◽  
R Saiki ◽  
TH Huisman ◽  
A Kutlar ◽  
V McKie ◽  
...  
Keyword(s):  
African American ◽  
African Americans ◽  
Ethnic Groups ◽  
Accurate Method ◽  
Beta Thalassemia ◽  
Dot Blot ◽  
Pcr Products ◽  
Hb S ◽  
Reverse Dot Blot ◽  
Hb C

DNA-based diagnosis of the beta thalassemias provides accuracy to newborn screening genetic counseling, and prenatal diagnosis. However, the use of polymerase chain reaction (PCR)-based methods is challenged by the great number of different-beta-thalassemia mutations that exist even within defined ethnic groups. In this regard, the reverse dot-blot method offers a means of screening for several mutations with a single hybridization reaction. We have applied the reverse dot-blot method to the detection of the beta-thalassemia mutations of African-Americans. We used two biotin-labeled primer pairs in a duplex reaction to amplify and label two beta-globin target DNA fragments that encompass all known African-American beta-thalassemia mutations. The PCR products were denatured and hybridized to polyT-tailed, membrane-fixed, allele- specific probe pairs for the hemoglobin (Hb) S, Hb C, and 14 beta- thalassemia mutations and their corresponding wild-type sequences. Seven common mutations plus Hb S and Hb C were included on one diagnostic strip, and seven less common beta-thalassemia mutations were included on another strip. Carefully controlled, high stringency hybridization allowed accurate distinction of these alleles. Reverse dot-blot diagnosis of the less common beta-thalassemia mutations precludes the need for alternative, more technically challenging methods. This method provides a rapid, accurate method for diagnosis of beta thalassemia among African-Americans and other ethnic groups in which beta thalassemia occurs.

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