A simple and rapid genotyping method for beta-2 receptor (beta2 AR) gene using allele specific multiplex PCR

2004 ◽  
Vol 29 (1) ◽  
pp. 47-52 ◽  
Author(s):  
S. M. N. Romaino ◽  
L. K. Teh ◽  
B. A. Zilfalil ◽  
C. P. Thong ◽  
A. A. Ismail ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genotyping Method ◽  
Allele Specific ◽  
Beta 2

Mycobacterium tuberculosis typing using Allele-specific oligonucleotide multiplex PCR (ASO–PCR) method

2021 ◽  
Author(s):  
Maryam Shafipour ◽  
Hesamaddin Shirzad-Aski ◽  
Ezzat Allah Ghaemi ◽  
Ahmad Sohrabi ◽  
Maya Babaii Kochaksaraei ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Multiplex Pcr ◽  
Allele Specific ◽  
Pcr Method

scholarly journals Direct detection of multiple point mutations in mitochondrial DNA

1997 ◽  
Vol 43 (10) ◽  
pp. 1857-1861 ◽  
Author(s):  
Lee-Jun C Wong ◽  
Dinithi Senadheera
Keyword(s):  
Mitochondrial Dna ◽  
Multiplex Pcr ◽  
Direct Detection ◽  
Point Mutations ◽  
Cost Effective ◽  
Large Deletion ◽  
Multiple Point ◽  
Mtdna Mutations ◽  
Allele Specific ◽  
Mitochondrial Defects

Abstract Mitochondrial defects can be caused by mutations in nuclear or mitochondrial DNA. Large deletion/duplication and point mutations are the two major types of mitochondrial DNA (mtDNA) mutations. Comprehensive molecular diagnosis requires the analysis of multiple point mutations. We developed an effective multiplex PCR/allele-specific oligonucleotide (ASO) method to simultaneously screen multiple point mutations in mtDNA. The system involved three pairs of primers to amplify mutation “hot spots” at tRNAleu(UUR), tRNAlys/ATPase, and ND4 regions, followed by detection of point mutations with ASO probes. Over 2000 specimens were analyzed and the results were compared with those from previous studies with the PCR/restriction fragment length polymorphism method. Our data demonstrate that the multiplex PCR/ASO method is much more sensitive in the detection of low mutant heteroplasmy. It is simple and cost effective, especially if a large number of samples are to be screened for multiple point mutations.

scholarly journals Improvement of Rotavirus Genotyping Method by Using the Semi-Nested Multiplex-PCR With New Primer Set

2019 ◽  
Vol 10 ◽  
Author(s):  
Yoshiki Fujii ◽  
Yen Hai Doan ◽  
Rury Mega Wahyuni ◽  
Maria Inge Lusida ◽  
Takako Utsumi ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genotyping Method

scholarly journals Microtransponder-Based Multiplex Assay for Genotyping Cystic Fibrosis

2007 ◽  
Vol 53 (7) ◽  
pp. 1372-1376 ◽  
Author(s):  
Xin Lin ◽  
James A Flint ◽  
Marco Azaro ◽  
Thomas Coradetti ◽  
Wesley M Kopacka ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
African American ◽  
Multiplex Pcr ◽  
False Positive ◽  
False Negative ◽  
Multiplex Assay ◽  
Specific Primer ◽  
Fluorescence Measurements ◽  
Allele Specific ◽  
Analysis Platform

Abstract Background: We developed and evaluated a genotyping assay for detection of 50 cystic fibrosis (CF) mutations. The assay is based on small (500 μm) electronic chips, radio frequency (RF) microtransponders (MTPs). The chips are analyzed on a unique fluorescence and RF readout instrument. Methods: We divided the CF assay into 4 panels: core, Hispanic, African-American, and Caucasian. We amplified 18 CF transmembrane regulator (CFTR) DNA fragments covering 50 mutations by use of multiplex PCR using 18 CFTR gene-specific primer pairs. PCR was followed by multiplex allele-specific primer extension (ASPE) reactions and hybridization to capture probes synthesized on MTPs. We used 100 ASPE primers and 100 capture probes. We performed fluorescence measurements of hybridized MTP kits and assay analysis using a custom automated bench-top flow instrument. Results: We validated the system by performing the assay on 23 commercial DNA samples in an internal study and 32 DNA samples in an external study. For internal and external studies, correct calls were 98.8% and 95.7%, false-positive calls 1.1% and 3.9%, and false-negative calls 0.12% and 0.36%, respectively. Conclusions: The MTP-based multiplex assay and analysis platform can be used for CF genotyping.

scholarly journals Multiplex PCR for simultaneous genotyping of kdr mutations V410L, V1,016I and F1,534C in Aedes aegypti (L.)

2019 ◽  
Author(s):  
Karina Villanueva-Segura ◽  
Gustavo Ponce-Garcia ◽  
Beatriz Lopez-Monroy ◽  
Esteban de J. Mora-Jasso ◽  
Lucia Perales ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Multiplex Pcr ◽  
Management Strategies ◽  
Simultaneous Detection ◽  
Resistance Management ◽  
Single Amino Acid ◽  
Specific Pcr ◽  
Highly Effective ◽  
Allele Specific ◽  
Current Monitoring

AbstractBackgroundKnock down resistance (kdr) is the main mechanism that confers resistance to pyrethroids and DDT. This is a product of nonsynonymous mutations in the he voltage-gated sodium channel (VGSC) gene, these mutations produce a change of a single amino acid which reduces the affinity of the target site for the insecticide molecule. In Mexico, V410L, V1,016I and F1,534C mutations are common in pyrethroid-resistant Aedes aegypti (L.) populations.MethodsA multiplex PCR was developed to detect the V410L, V1,016I and F1,534C mutations in Ae. aegypti. The validation of the technique was carried out using wild populations previously characterized for the three mutations through allele-specific PCR (AS-PCR) and with different levels of genotypic frequencies.ResultsThe standardized protocol for multiplex endpoint PCR was highly effective in detecting 12 genotypes in five wild Ae. aegypti populations from Mexico. A complete concordance with AS-PCR was found for the simultaneous detection of the three kdr mutations.ConclusionsOur diagnostic method is highly effective for the simultaneous detection of V410L, V1,016I and F1,534C, when they co-occur. This technique represents a viable alternative to complement and strengthen current monitoring and resistance management strategies against Ae. aegypti.

scholarly journals A Rapid Robust Method for Subgrouping Non-NF2 Meningiomas According to Genotype and Detection of Lower Levels of M2 Macrophages in AKT1 E17K Mutated Tumours

2020 ◽  
Vol 21 (4) ◽  
pp. 1273 ◽  
Author(s):  
Claire L. Adams ◽  
Emanuela Ercolano ◽  
Sara Ferluga ◽  
Agbolahan Sofela ◽  
Foram Dave ◽  
...  
Keyword(s):  
Immune Cell ◽  
Tumour Microenvironment ◽  
M2 Macrophages ◽  
Who Grade ◽  
Specific Pcr ◽  
Genotyping Method ◽  
Mutational Status ◽  
Hla Dr ◽  
Allele Specific ◽  
Allele Specific Pcr

The majority of meningiomas are grade I, but some grade I tumours are clinically more aggressive. Recent advances in the genetic study of meningiomas has allowed investigation into the influence of genetics on the tumour microenvironment, which is important for tumorigenesis. We have established that the endpoint genotyping method Kompetitive Allele Specific PCR (KASP™) is a fast, reliable method for the screening of meningioma samples into different non-NF2 mutational groups using a standard real-time PCR instrument. This genotyping method and four-colour flow cytometry has enabled us to assess the variability in the largest immune cell infiltrate population, M2 macrophages (CD45+HLA-DR+CD14+CD163+) in 42 meningioma samples, and to suggest that underlying genetics is relevant. Further immunohistochemistry analysis comparing AKT1 E17K mutants to WHO grade I NF2-negative samples showed significantly lower levels of CD163-positive activated M2 macrophages in meningiomas with mutated AKT1 E17K, signifying a more immunosuppressive tumour microenvironment in NF2 meningiomas. Our data suggested that underlying tumour genetics play a part in the development of the immune composition of the tumour microenvironment. Stratifying meningiomas by mutational status and correlating this with their cellular composition will aid in the development of new immunotherapies for patients.

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