Heteroplasmy Detection of Mitochondrial DNA A3243G Mutation Using Quantitative Real-Time PCR Assay Based on TaqMan-MGB Probes

المؤلفون المشاركون

Rong, Enguang
Wang, Hanbo
Hao, Shujing
Fu, Yuhong
Ma, Yanyan
Wang, Tianze

المصدر

BioMed Research International

العدد

المجلد 2018، العدد 2018 (31 ديسمبر/كانون الأول 2018)، ص ص. 1-9، 9ص.

الناشر

Hindawi Publishing Corporation

تاريخ النشر

2018-11-13

دولة النشر

مصر

عدد الصفحات

9

التخصصات الرئيسية

الطب البشري

الملخص EN

A point mutation of mitochondrial DNA (mtDNA) at nucleotide position 3243 A to G (mt.3243A>G) is involved in many common diseases, including maternally inherited diabetes and deafness (MIDD) and mitochondrial encephalomyopathy, lactic acidosis with stroke-like episodes (MELAS).

However, the mutant level of mt.3243A>G varies both among individuals and in different organs, tissues, and even cells of single individuals.

For detection of this mutation, current methods have limited universality and sensitivity and may be not adequate for a routine clinical test.

Here, we develop and evaluate a rapid TaqMan-MGB quantitative real-time PCR (qPCR) method for detecting and quantifying the heteroplasmy level of mt.3243A>G in single-tube analysis.

With our method, the sensitivity of detection was as low as 0.1%, but the accuracy of quantification was reliable, down to 4%.

All positives could be correctly identified, and the heteroplasmy levels determined by qPCR correlated well with the results from restriction fragment length polymorphism (RFLP) and pyrosequencing assays (r = 0.921~0.973 and 0.972~0.984).

In addition, we demonstrated that the urinary sediments, leukocytes, or hair follicles might be ideal templates to detect and quantify the heteroplasmy of mt.3243A>G mutation; however, they should be optimized or retreated for further accurate quantification.

Our study should allow rapid and high throughput diagnostic testing and can potentially be used to clarify the association between clinical phenotype and pathogenic mitochondrial mutations derived from various tissues.

نمط استشهاد جمعية علماء النفس الأمريكية (APA)

Rong, Enguang& Wang, Hanbo& Hao, Shujing& Fu, Yuhong& Ma, Yanyan& Wang, Tianze. 2018. Heteroplasmy Detection of Mitochondrial DNA A3243G Mutation Using Quantitative Real-Time PCR Assay Based on TaqMan-MGB Probes. BioMed Research International،Vol. 2018, no. 2018, pp.1-9.
https://search.emarefa.net/detail/BIM-1124215

نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)

Rong, Enguang…[et al.]. Heteroplasmy Detection of Mitochondrial DNA A3243G Mutation Using Quantitative Real-Time PCR Assay Based on TaqMan-MGB Probes. BioMed Research International No. 2018 (2018), pp.1-9.
https://search.emarefa.net/detail/BIM-1124215

نمط استشهاد الجمعية الطبية الأمريكية (AMA)

Rong, Enguang& Wang, Hanbo& Hao, Shujing& Fu, Yuhong& Ma, Yanyan& Wang, Tianze. Heteroplasmy Detection of Mitochondrial DNA A3243G Mutation Using Quantitative Real-Time PCR Assay Based on TaqMan-MGB Probes. BioMed Research International. 2018. Vol. 2018, no. 2018, pp.1-9.
https://search.emarefa.net/detail/BIM-1124215

نوع البيانات

مقالات

لغة النص

الإنجليزية

الملاحظات

Includes bibliographical references

رقم السجل

BIM-1124215