Development of a Multiplex PCR for Human Neutrophil Antigens-1, -3, -4, and -5 Genotyping Among Thai Blood Donors

Nathalang, Oytip; Intharanut, Kamphon; Siriphanthong, Kanokpol; Sasikarn, Wiradee; Nathalang, Siriporn

doi:10.7754/clin.lab.2016.160432

Cited by 3 publications

(6 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nevertheless, individual testing for each antigen may not be suitable to find antigen-negative blood units. The implementation of multiplex-PCR is beneficial for transfusion-dependent patients for the following reasons: i) The results can be obtained even with a low concentration of DNA (20 ng/μl) within 2 h. ii) Multiplex PCR uses fewer tubes and less reagents, and the cost is approximately Eur 4.00/test, which is 50% less than PCR-SSP, and similar to the application of multiplex PCR to detect other genetic polymorphisms [30,31]. iii) Its simplicity and reproducibility is proper for routine testing.…”

Section: Discussionmentioning

confidence: 99%

Red Cell Genotyping by Multiplex PCR Identifies Antigen-Matched Blood Units for Transfusion-Dependent Thai Patients

Intharanut

Bejrachandra

Nathalang

et al. 2017

Transfus Med Hemother

Self Cite

View full text Add to dashboard Cite

Background: Antigen-negative red cell transfusion is required for transfusion-dependent patients. We developed multiplex PCR for red cell genotyping and calculated the possibility of finding compatible predicted phenotypes in Thai blood donor populations according to red cell alloantibodies found among Thai patients. Methods: 600 DNA samples obtained from unrelated healthy central and northern Thai blood donors were tested with the newly developed multiplex PCR for FY*A, FY*B, JK*A, JK*B, RHCE*e, RHCE*E, DI*A and GYP*Hut, GYP*Mur, GYP*Hop, GYP*Bun, and GYP*HF allele detections. Additionally, the possibility of finding compatible predicted phenotypes in two Thai blood donor populations was calculated to estimate the minimal number of tests needed to provide compatible blood. Results: The validity of multiplex PCR using known DNA controls and the phenotyping and genotyping results obtained by serological and PCR-SSP techniques were in agreement. The possibility of finding at least one compatible blood unit for patients with multiple antibodies was comparable in Thai populations. Conclusions: The multiplex PCR for red cell genotyping simultaneously interprets 7 alleles and 1 hybrid GP group. Similar strategies can be applied in other populations depending on alloantibody frequencies in transfusion-dependent patients, especially in a country with limited resources.

show abstract

Section: Discussionmentioning

confidence: 99%

Red Cell Genotyping by Multiplex PCR Identifies Antigen-Matched Blood Units for Transfusion-Dependent Thai Patients

Intharanut

Bejrachandra

Nathalang

et al. 2017

Transfus Med Hemother

Self Cite

View full text Add to dashboard Cite

show abstract

“…HNA‐1, ‐3, ‐4, and ‐5 genotyping, using multiplex PCR, was performed as previously described . Briefly, the multiplex PCR for HNA‐1a, ‐1b, ‐1c, ‐3a, ‐3b, ‐4a, ‐4b, ‐5a, and ‐5b genotyping contained four multiple reaction sets (A to D) of each different amplification target per mix.…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, only HNA‐4 genotype frequencies among northeastern Thais were similar to central Thais . To determine more convenient HNA genotyping, multiplex PCR has been applied to reduce costs and time‐consuming tasks by performing the simultaneous detection of HNA‐1, ‐3, ‐4, and ‐5 . The data of HNA among southern Thais remain unknown.…”

Section: Introductionmentioning

confidence: 99%

“…Several cases of neonatal neutropenia caused by HNA‐1 antibodies of FCGR3B‐ deficient mothers were reported in different populations . Regarding related studies, genotyping of the HNA‐1 null individuals using PCR‐SSP and/or multiplex PCR showed no bands of FCGR3B*01 , FCGR3B*02 , and FCGR3B*03 allele detections . The PCR‐restriction fragment length polymorphism (RFLP) is suggested to be used to confirm FCGR3B‐ deficient individuals .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

HNA‐1, ‐3, ‐4, and ‐5 genotyping using multiplex PCR among southern Thais: Developing continual HNA‐1 null detection

Intharanut

Sasikarn

Mitundee

et al. 2018

Clinical Laboratory Analysis

Self Cite

View full text Add to dashboard Cite

show abstract

“…By combining 2D PCR with other gene detection technologies, the range of detectable genes can be expanded. For instance, we can combine 2D PCR with the PCR-specific sequence primer method (PCR-SSP) to detect the mutation of different genes by adding tags to the specific sequence primers. , …”

Section: Resultsmentioning

confidence: 99%

High-Throughput Two-Dimensional Polymerase Chain Reaction Technology

et al. 2019

View full text Add to dashboard Cite

Polymerase chain reaction (PCR) is a very powerful tool for clinical gene detection. Multiplex PCR especially improves the throughput of this technology. However, it is often necessary to employ techniques such as electrophoresis, mass spectrometry, or sequencing after multiplex PCR amplification for product identification, which requires additional equipment and has high risks of contamination. In this work, we developed a high-throughput two-dimensional (2D) PCR technology that can identify multiple target genes simultaneously in just one closed tube and within a relatively short time by using both fluorescence and the melting temperature (Tm). As an example, a method detecting 9 human papillomavirus (HPV) subtypes and reference genes in a single tube was successfully established using 2D PCR. If designed properly, 2D PCR is believed to have the capability to identify more than 30 genes in one closed tube at a time. This method is particularly suitable for distinguishing microorganisms, single-nucleotide polymorphisms, and the methylation of genes and will be of great help to clinical work.

show abstract

Development of a Multiplex PCR for Human Neutrophil Antigens-1, -3, -4, and -5 Genotyping Among Thai Blood Donors

Cited by 3 publications

References 0 publications

Red Cell Genotyping by Multiplex PCR Identifies Antigen-Matched Blood Units for Transfusion-Dependent Thai Patients

Red Cell Genotyping by Multiplex PCR Identifies Antigen-Matched Blood Units for Transfusion-Dependent Thai Patients

HNA‐1, ‐3, ‐4, and ‐5 genotyping using multiplex PCR among southern Thais: Developing continual HNA‐1 null detection

High-Throughput Two-Dimensional Polymerase Chain Reaction Technology

Contact Info

Product

Resources

About