Correction to ‘Genotyping of Malaysian G6PD-deficient neonates by reverse dot blot flow-through hybridisation’

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

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Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes

BioMed Research International ◽

10.1155/2014/156323 ◽

2014 ◽

Vol 2014 ◽

pp. 1-11 ◽

Cited By ~ 7

Author(s):

K. Böhme ◽

P. Cremonesi ◽

M. Severgnini ◽

Tomás G. Villa ◽

I. C. Fernández-No ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Bacterial Species ◽

Cost Effective ◽

Rrna Gene ◽

Bacterial Identification ◽

Dot Blot ◽

Culture Collections ◽

Ligation Detection Reaction ◽

Flow Through ◽

Food Safety And Quality

Traditional culturing methods are still commonly applied for bacterial identification in the food control sector, despite being time and labor intensive. Microarray technologies represent an interesting alternative. However, they require higher costs and technical expertise, making them still inappropriate for microbial routine analysis. The present study describes the development of an efficient method for bacterial identification based on flow-through reverse dot-blot (FT-RDB) hybridization on membranes, coupled to the high specific ligation detection reaction (LDR). First, the methodology was optimized by testing different types of ligase enzymes, labeling, and membranes. Furthermore, specific oligonucleotide probes were designed based on the 16S rRNA gene, using the bioinformatic tool Oligonucleotide Retrieving for Molecular Applications (ORMA). Four probes were selected and synthesized, being specific forAeromonasspp.,Pseudomonasspp.,Shewanellaspp., andMorganella morganii, respectively. For the validation of the probes, 16 reference strains from type culture collections were tested by LDR and FT-RDB hybridization using universal arrays spotted onto membranes. In conclusion, the described methodology could be applied for the rapid, accurate, and cost-effective identification of bacterial species, exhibiting special relevance in food safety and quality.

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Simplified reverse dot blot analyses for detecting ras oncogene mutations

Molecular Diagnosis ◽

10.1016/s1084-8592(97)80025-2 ◽

1997 ◽

Vol 2 (3) ◽

pp. 169-176 ◽

Cited By ~ 12

Author(s):

M ALBITAR ◽

W WU ◽

E FELTZ ◽

G JIN ◽

C HIRSCHGINSBERG ◽

...

Keyword(s):

Ras Oncogene ◽

Dot Blot ◽

Reverse Dot Blot

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Hybridoma Screening by Antigen Capture: Reverse Dot Blot

Cold Spring Harbor Protocols ◽

10.1101/pdb.prot103135 ◽

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).

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Alpha and beta-Thalassemia mutations in Hubei area of China

BMC Medical Genetics ◽

10.1186/s12881-019-0925-5 ◽

2020 ◽

Vol 21 (1) ◽

Cited By ~ 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.

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