Home Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations
Article
Licensed
Unlicensed Requires Authentication

Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations

  • Cia-Hin Lau ORCID logo , Kejiang Guo , Gang Chen , Minghai Zou , Zhongqi Zhou , Tao Wang , Zhihao Huang , Jiaqi Li , Wenjiao Dong , Yumei Huang , Pik Kwan Lo , Hongman Xue , Xiaojun Huang , Meijing Xu , Chung Tin EMAIL logo and Haibao Zhu EMAIL logo
Published/Copyright: March 17, 2025
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 63 Issue 7

Abstract

Objectives

Detecting point mutations with high sensitivity and specificity can be technically very challenging, but it is crucial for early diagnosis and effective drug treatment of cancers. To enable ultrasensitive and ultraspecific detection of single-base mutations in simple and economical ways, we have developed an artificial base mismatches-mediated PCR (ABM-PCR) detection approach.

Methods

ABM-PCR was applied to quantitative PCR (qPCR) and droplet digital PCR (ddPCR) detection platforms. The impact of mismatches on the thermodynamic stability of the primer-template duplex and the ability of Taq polymerase to catalyze the extension was examined. Effects of the sequence, position, and the number of mismatches on genotyping performance were characterized.

Results

As proof of principle, we demonstrated the feasibility of ABM-PCR in detecting epidermal growth factor receptor (EGFR) and B-Raf proto-oncogene, serine/threonine kinase (BRAF) mutations that are clinically relevant to diagnosis and prognosis of lung and thyroid cancers. Our ABM-PCR enabled the detection of 0.1 % mutation without amplification of the wild-type DNA strand, even in the presence of a 300 ng human genomic DNA background. It enables ultrasensitive (≥95 %) and ultraspecific (≥95 %) diagnosis of clinical samples for thyroid papilloma and lung cancers. Based on these findings, we have established a set of rules and developed a user-friendly web primer design tool for designing effective ABM-PCR primers.

Conclusions

This study highlights the impact of primer-template mismatches on PCR amplification and provides insights into rational design of effective ABM-PCR primers for detecting single-base mutations with high specificity and sensitivity. It is highly valuable for clinical diagnosis and prognosis use.

Keywords: ARMS-PCR; base substitution; point mutation; polymerase extension; SNP; thermodynamic stability
Corresponding authors: Professor Chung Tin, Department of Biomedical Engineering, City University of Hong Kong, Hong Kong, China, E-mail: ; and Professor Haibao Zhu, Department of Biology, College of Science, Shantou, Guangdong, China; Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou, Guangdong, China; and Shantou Key Laboratory of Marine Microbial Resources and Interactions with Environment, 12386 Shantou University , Shantou, Guangdong, China, E-mail:
Cia-Hin Lau, Kejiang Guo, Gang Chen and Minghai Zou contributed equally to this work.

Funding source: Shantou University Research Initiation Fund Project

Award Identifier / Grant number: NTF20030

Funding source: Guangdong Provincial Natural Science Foundation General Project

Award Identifier / Grant number: 2023A1515011906

Funding source: City University of Hong Kong

Award Identifier / Grant number: 7020051

Funding source: Sanming Project of Medicine in Shenzhen

Award Identifier / Grant number: SZSM202011004

Funding source: Shenzhen Science and Technology Innovation Commission

Award Identifier / Grant number: JCYJ20190809160609727

Funding source: Xiamen Municipal Bureau of Science and Technology-National Foreign Expert Program

Award Identifier / Grant number: QN2023021001L

  1. Research ethics: The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). Ethical clearance to undertake this study was obtained from the Fujian Cancer Hospital Ethics Committee (reference number of Ethical Approval Letter was SQ2024-176).

  2. Informed consent: Not applicable.

  3. Author contributions: H.Z, C.T., and C.H.L. conceived the project and designed the experiments. C.H.L., K.G., G.C., Z.Z., T.W., Z.H., J.L., W.D., Y.H., P.K.L., H.X., and X.H. performed the experiments, collected and analyzed the data. H.Z, C.T., and C.H.L. wrote and revised the manuscript with help from all authors. All authors read, corrected, and approved the final manuscript.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflicts of interest.

  6. Research funding: This work was supported by Shantou University Research Initiation Fund Project (NTF20030), Guangdong Provincial Natural Science Foundation General Project (2023A1515011906), Shenzhen Science and Technology Innovation Commission (JCYJ20190809160609727), Sanming Project of Medicine in Shenzhen (SZSM202011004), Xiamen Municipal Bureau of Science and Technology-National Foreign Expert Program (QN2023021001L), City University of Hong Kong (7020051).

  7. Data availability: The datasets generated and/or analyzed during this study are included in this article (and its supplementary material file). Any additional data, if needed, will be made on reasonable request to the corresponding author.

References

1. He, C, Wei, C, Wen, J, Chen, S, Chen, L, Wu, Y, et al.. Comprehensive analysis of NGS and ARMS-PCR for detecting EGFR mutations based on 4467 cases of NSCLC patients. J Cancer Res Clin Oncol 2022;148:321–30. https://doi.org/10.1007/s00432-021-03818-w.Search in Google Scholar PubMed PubMed Central

2. Zhou, C, Ni, J, Zhao, Y, Su, B. Rapid detection of epidermal growth factor receptor mutations in non-small cell lung cancer using real-time polymerase chain reaction with TaqMan-MGB probes. Cancer J 2006;12:33–9. https://doi.org/10.1097/00130404-200601000-00007.Search in Google Scholar PubMed

3. Ayyadevara, S, Thaden, JJ, Shmookler Reis, RJ. Discrimination of primer 3’-nucleotide mismatch by taq DNA polymerase during polymerase chain reaction. Anal Biochem 2000;284:11–18. https://doi.org/10.1006/abio.2000.4635.Search in Google Scholar PubMed

4. Huang, K, Zhang, J, Li, J, Qiu, H, Wei, L, Yang, Y, et al.. Exploring the impact of primer-template mismatches on PCR performance of DNA polymerases varying in proofreading activity. Genes 2024;15:215. https://doi.org/10.3390/genes15020215.Search in Google Scholar PubMed PubMed Central

5. Huang, MM, Arnheim, N, Goodman, MF. Extension of base mispairs by Taq DNA polymerase: implications for single nucleotide discrimination in PCR. Nucleic Acids Res 1992;20:4567–73. https://doi.org/10.1093/nar/20.17.4567.Search in Google Scholar PubMed PubMed Central

6. Chen, M, Huang, W, Yang, D, Huang, J, Li, G, Wang, X, et al.. DeltaCT value of amplified refractory mutation system predicts efficacy of EGFR-TKIs in advanced non-small-cell lung cancer: a multi-center retrospective study. Front Mol Biosci 2021;8:684661. https://doi.org/10.3389/fmolb.2021.684661.Search in Google Scholar PubMed PubMed Central

7. Roma, C, Esposito, C, Rachiglio, AM, Pasquale, R, Iannaccone, A, Chicchinelli, N, et al.. Detection of EGFR mutations by TaqMan mutation detection assays powered by competitive allele-specific TaqMan PCR technology. BioMed Res Int 2013;2013:385087. https://doi.org/10.1155/2013/385087.Search in Google Scholar PubMed PubMed Central

8. Sun, Y, Yin, X, Wen, MM, Zhang, J, Wang, XJ, Xia, JH, et al.. EGFR mutations subset in Chinese lung squamous cell carcinoma patients. Mol Med Rep 2018;17:7575–84. https://doi.org/10.3892/mmr.2018.8859.Search in Google Scholar PubMed PubMed Central

9. Zhang, X, Chang, N, Yang, G, Zhang, Y, Ye, M, Cao, J, et al.. A comparison of ARMS-Plus and droplet digital PCR for detecting EGFR activating mutations in plasma. Oncotarget 2017;8:112014–23. https://doi.org/10.18632/oncotarget.22997.Search in Google Scholar PubMed PubMed Central

10. Liu, Q, Sommer, SS. PAP: detection of ultra rare mutations depends on P* oligonucleotides: “sleeping beauties” awakened by the kiss of pyrophosphorolysis. Hum Mutat 2004;23:426–36. https://doi.org/10.1002/humu.20036.Search in Google Scholar PubMed

11. Chen, Y, Zhao, X, Wang, L, Wu, F, Zhang, X, Wu, H, et al.. Super-ARMS: a new method for plasma ESR1 mutation detection. Clin Chim Acta 2021;520:23–8. https://doi.org/10.1016/j.cca.2021.05.021.Search in Google Scholar PubMed

12. Xu, G, Yang, H, Qiu, J, Reboud, J, Zhen, L, Ren, W, et al.. Sequence terminus dependent PCR for site-specific mutation and modification detection. Nat Commun 2023;14:1169. https://doi.org/10.1038/s41467-023-36884-4.Search in Google Scholar PubMed PubMed Central

13. Zhang, Z, Shan, X, Zhang, P, Liu, W, Yan, J, Li, Z. Highly sensitive quantification of site-specific 5-hydroxymethylcytosine at single-base resolution by HpaII-mediated ligation PCR. Org Biomol Chem 2019;17:9849–53. https://doi.org/10.1039/c9ob02429h.Search in Google Scholar PubMed

14. Deng, P, Liu, X, Li, Y, Li, H, Zhao, B, Wang, S, et al.. Standardization and application of ARMS TaqMan real-time PCR for screening of folate metabolism genes in Han Chinese. Electrophoresis 2024;45:1995–2004. https://doi.org/10.1002/elps.202400017.Search in Google Scholar PubMed

15. Nie, H, Evans, AA, London, WT, Block, TM, Ren, XD. Ultrasensitive quantification of hepatitis B virus A1762T/G1764A mutant by a SimpleProbe PCR using a wild-type-selective PCR blocker and a primer-blocker-probe partial-overlap approach. J Clin Microbiol 2011;49:2440–8. https://doi.org/10.1128/jcm.02472-10.Search in Google Scholar

16. Huang, Q, Wang, GY, Huang, JF, Zhang, B, Fu, WL. High sensitive mutation analysis on KRAS gene using LNA/DNA chimeras as PCR amplification blockers of wild-type alleles. Mol Cell Probes 2010;24:376–80. https://doi.org/10.1016/j.mcp.2010.07.010.Search in Google Scholar PubMed

17. Xie, F, Huang, J, Qu, S, Wu, W, Jiang, J, Wang, H, et al.. Sensitive detection of trace amounts of KRAS codon 12 mutations by a fast and novel one-step technique. Clin Biochem 2014;47:237–42. https://doi.org/10.1016/j.clinbiochem.2014.08.015.Search in Google Scholar PubMed

18. Chubarov, AS, Baranovskaya, EE, Oscorbin, IP, Yushin, II, Filipenko, ML, Pyshnyi, DV, et al.. Phosphoramidate azole oligonucleotides for single nucleotide polymorphism detection by PCR. Int J Mol Sci 2024;25:617. https://doi.org/10.3390/ijms25010617.Search in Google Scholar PubMed PubMed Central

19. Fujita, H, Fujita, T, Ishido, K, Hakamada, K, Fujii, H. Oligoribonucleotide interference-PCR-based methods for the sensitive and accurate detection of KRAS mutations. Biol Methods Protoc 2024;9:bpae071. https://doi.org/10.1093/biomethods/bpae071.Search in Google Scholar PubMed PubMed Central

20. Nam, H, Lee, E, Yang, H, Lee, K, Kwak, T, Kim, D, et al.. PROMER technology: a new real-time PCR tool enabling multiplex detection of point mutation with high specificity and sensitivity. Biol Methods Protoc 2024;9:bpae041. https://doi.org/10.1093/biomethods/bpae041.Search in Google Scholar PubMed PubMed Central

21. Liu, Y, Liu, Y, Guo, L, Wu, Y, Wang, Y, Xu, L, et al.. Multiplex asymmetric PCR by combining the amplification refractory mutation system with the homo-tag-assisted nondimer system. Anal Chem 2024;96:9200–8. https://doi.org/10.1021/acs.analchem.4c01322.Search in Google Scholar PubMed

22. Debouki-Joudi, S, Ben Kridis, W, Trifa, F, Ayadi, W, Khabir, A, Sellami-Boudawara, T, et al.. A novel PIK3CA hot-spot mutation in breast cancer patients detected by HRM-COLD-PCR analysis. Breast Dis 2024;43:213–21. https://doi.org/10.3233/bd-240005.Search in Google Scholar PubMed PubMed Central

23. Mirikar, D, Banerjee, N, Prabhash, K, Kaushal, RK, Naronha, V, Pramesh, CS, et al.. Comparative analysis of EGFR mutations in circulating tumor DNA and primary tumor tissues from lung cancer patients using BEAMing PCR. Sci Rep 2025;15:1252. https://doi.org/10.1038/s41598-025-85160-6.Search in Google Scholar PubMed PubMed Central

24. Wu, JH, Hong, PY, Liu, WT. Quantitative effects of position and type of single mismatch on single base primer extension. J Microbiol Methods 2009;77:267–75. https://doi.org/10.1016/j.mimet.2009.03.001.Search in Google Scholar PubMed

25. Kwok, S, Kellogg, DE, McKinney, N, Spasic, D, Goda, L, Levenson, C, et al.. Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies. Nucleic Acids Res 1990;18:999–1005. https://doi.org/10.1093/nar/18.4.999.Search in Google Scholar PubMed PubMed Central

26. Kaltenbrunner, M, Hochegger, R, Cichna-Markl, M. Design of mismatch primers to identify and differentiate closely related (sub) species: application to the authentication of meat products. Methods Mol Biol 2022;2392:65–82. https://doi.org/10.1007/978-1-0716-1799-1_5.Search in Google Scholar PubMed

27. Lefever, S, Rihani, A, Van Der Meulen, J, Pattyn, F, Van Maerken, T, Van Dorpe, J, et al.. Cost-effective and robust genotyping using double-mismatch allele-specific quantitative PCR. Sci Rep 2019;9:2150. https://doi.org/10.1038/s41598-019-38581-z.Search in Google Scholar PubMed PubMed Central

28. Thomas, BJ, Guldenpfennig, C, Guan, Y, Winkler, C, Beecher, M, Beedy, M, et al.. Targeting lung cancer with clinically relevant EGFR mutations using anti-EGFR RNA aptamer. Mol Ther Nucleic Acids 2023;34:102046. https://doi.org/10.1016/j.omtn.2023.102046.Search in Google Scholar PubMed PubMed Central

29. Kitamura, A, Ishii, K, Okafuji, K, Kojima, F, Bando, T. Epidermal growth factor receptor (EGFR) mutation testing is useful for primary lung cancer diagnosis and appropriate surgical resection: a case series. Respir Investig 2022;60:171–5. https://doi.org/10.1016/j.resinv.2021.08.008.Search in Google Scholar PubMed

30. De Biase, D, Visani, M, Malapelle, U, Simonato, F, Cesari, V, Bellevicine, C, et al.. Next-generation sequencing of lung cancer EGFR exons 18-21 allows effective molecular diagnosis of small routine samples (cytology and biopsy). PLoS One 2013;8:e83607. https://doi.org/10.1371/journal.pone.0083607.Search in Google Scholar PubMed PubMed Central

31. Bordi, P, Del, RM, Danesi, R, Tiseo, M. Circulating DNA in diagnosis and monitoring EGFR gene mutations in advanced non-small cell lung cancer. Transl Lung Cancer Res 2015;4:584–97. https://doi.org/10.3978/j.issn.2218-6751.2015.08.09.Search in Google Scholar PubMed PubMed Central

32. Adeniran, AJ, Hui, P. Best practice of BRAF V600E mutation testing for the diagnosis and management of thyroid cancers. Expet Rev Endocrinol Metabol 2014;9:571–7. https://doi.org/10.1586/17446651.2014.951635.Search in Google Scholar PubMed

33. Trovisco, V, Soares, P, Sobrinho-Simoes, M. B-RAF mutations in the etiopathogenesis, diagnosis, and prognosis of thyroid carcinomas. Hum Pathol 2006;37:781–6. https://doi.org/10.1016/j.humpath.2006.03.013.Search in Google Scholar PubMed

34. Wang, Z, Chen, JQ, Liu, JL, Qin, XG. Clinical impact of BRAF mutation on the diagnosis and prognosis of papillary thyroid carcinoma: a systematic review and meta-analysis. Eur J Clin Invest 2016;46:146–57. https://doi.org/10.1111/eci.12577.Search in Google Scholar PubMed

35. Yu, PC, Tan, LC, Zhu, XL, Shi, X, Chernikov, R, Semenov, A, et al.. Arms-qPCR improves detection sensitivity of earlier diagnosis of papillary thyroid cancers with worse prognosis determined by coexisting BRAF V600E and Tert promoter mutations. Endocr Pract 2021;27:698–705. https://doi.org/10.1016/j.eprac.2021.01.015.Search in Google Scholar PubMed

36. Stadhouders, R, Pas, SD, Anber, J, Voermans, J, Mes, TH, Schutten, M. The effect of primer-template mismatches on the detection and quantification of nucleic acids using the 5’ nuclease assay. J Mol Diagn 2010;12:109–17. https://doi.org/10.2353/jmoldx.2010.090035.Search in Google Scholar PubMed PubMed Central

37. Johnson, SJ, Beese, LS. Structures of mismatch replication errors observed in a DNA polymerase. Cell 2004;116:803–16. https://doi.org/10.1016/s0092-8674(04)00252-1.Search in Google Scholar PubMed

38. Rejali, NA, Moric, E, Wittwer, CT. The effect of single mismatches on primer extension. Clin Chem 2018;64:801–9. https://doi.org/10.1373/clinchem.2017.282285.Search in Google Scholar PubMed

39. Zhang, BO, Xu, CW, Shao, Y, Wang, HT, Wu, YF, Song, YY, et al.. Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation. Exp Ther Med 2015;9:1383–8. https://doi.org/10.3892/etm.2015.2221.Search in Google Scholar PubMed PubMed Central

40. Bizouarn, F. Clinical applications using digital PCR. Methods Mol Biol 2014;1160:189–214. https://doi.org/10.1007/978-1-4939-0733-5_16.Search in Google Scholar PubMed

41. Bhatia, P, Thakur, R, Sreedharanunni, S, Singh, M, Malhotra, M, Arora, S, et al.. Clinical utility of a novel triplex digital PCR assay for clone monitoring in sequential and relapsed pediatric B-cell acute lymphoblastic leukemia patients. Pediatr Blood Cancer 2025;72:e31443. https://doi.org/10.1002/pbc.31443.Search in Google Scholar PubMed

42. Aran, V, Lamback, E, Lyra Miranda, R, Guterres, A, Souza Barbosa, I, Chimelli, L, et al.. Using digital PCR to investigate the prevalence of KRAS variants in pituitary tumours. J Neuroendocrinol 2025;37:e13484. https://doi.org/10.1111/jne.13484.Search in Google Scholar PubMed

43. Zhong, Y, Tan, GW, Bult, J, Veltmaat, N, Plattel, W, Kluiver, J, et al.. Detection of circulating tumor DNA in plasma of patients with primary CNS lymphoma by digital droplet PCR. BMC Cancer 2024;24:407. https://doi.org/10.1186/s12885-024-12191-z.Search in Google Scholar PubMed PubMed Central

44. Wu, S, Li, Y, Huang, R, Li, T, Yu, Y, Luo, P, et al.. Detection and quantification of JAK2V617F copy number by droplet digital PCR versus real-time PCR. Ann Hematol 2024;103:421–6. https://doi.org/10.1007/s00277-023-05544-4.Search in Google Scholar PubMed

45. Sanchez-Martin, V, Lopez-Lopez, E, Reguero-Paredes, D, Godoy-Ortiz, A, Dominguez-Recio, ME, Jimenez-Rodriguez, B, et al.. Comparative study of droplet-digital PCR and absolute Q digital PCR for ctDNA detection in early-stage breast cancer patients. Clin Chim Acta 2024;552:117673. https://doi.org/10.1016/j.cca.2023.117673.Search in Google Scholar PubMed

46. Min, YK, Kim, JK, Park, KS, Kim, JW. Evaluation of droplet digital PCR for the detection of BRAF V600E in fine-needle aspiration specimens of thyroid nodules. Ann Lab Med 2024;44:553–61. https://doi.org/10.3343/alm.2023.0405.Search in Google Scholar PubMed PubMed Central

47. Liu, Y, Han, C, Li, J, Xu, S, Xiao, Z, Guo, Z, et al.. Laboratory-developed droplet digital PCR assay for quantification of the JAK2 (V617F) mutation. Glob Med Genet 2024;11:132–41. https://doi.org/10.1055/s-0044-1785537.Search in Google Scholar PubMed PubMed Central

48. Hu, L, Ji, YY, Zhu, P, Lu, RQ. Mutation-Selected Amplification droplet digital PCR: a new single nucleotide variant detection assay for TP53(R249S) mutant in tumor and plasma samples. Anal Chim Acta 2024;1318:342929. https://doi.org/10.1016/j.aca.2024.342929.Search in Google Scholar PubMed

49. Zhang, Z, Li, L, Yao, J. Multiplex one-step direct asymmetric PCR of blood and dual-labelled probe-mediated melting curve for genotyping of MTHFR and MTRR polymorphisms. RSC Adv 2025;15:75–82. https://doi.org/10.1039/d4ra07286c.Search in Google Scholar PubMed PubMed Central

50. Zhang, F, Liu, ZY, Liu, S, Zhang, WG, Wang, BB, Li, CL, et al.. Rapid screening of point mutations by mismatch amplification mutation assay PCR. Appl Microbiol Biotechnol 2024;108:190. https://doi.org/10.1007/s00253-024-13036-2.Search in Google Scholar PubMed PubMed Central

51. Yao, H, Wang, K, Lu, S, Cao, F, Dai, P. Development of an ARMS multiplex real-time PCR assay for the detection of HLA-B*13:01 genotype by detecting highly specific SNPs. Pharmacogenet Genomics 2024;34:53–9. https://doi.org/10.1097/FPC.0000000000000517.Search in Google Scholar PubMed

52. Gullu, AG, Polat, B, Kahraman, A, Akkiprik, M. A simple and cost-effective real-time PCR method using diluted and heat-treated whole blood lysate. Sci Rep 2024;14:27225. https://doi.org/10.1038/s41598-024-78802-8.Search in Google Scholar PubMed PubMed Central

Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2024-0962).

Received: 2024-08-19
Accepted: 2025-02-23
Published Online: 2025-03-17
Published in Print: 2025-06-26

© 2025 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. The journey to pre-analytical quality
  4. Manual tilt tube method for prothrombin time: a commentary on contemporary relevance
  5. Reviews
  6. From errors to excellence: the pre-analytical journey to improved quality in diagnostics. A scoping review
  7. Advancements and challenges in high-sensitivity cardiac troponin assays: diagnostic, pathophysiological, and clinical perspectives
  8. Opinion Paper
  9. Is it feasible for European laboratories to use SI units in reporting results?
  10. Perspectives
  11. What does cancer screening have to do with tomato growing?
  12. Computer simulation approaches to evaluate the interaction between analytical performance characteristics and clinical (mis)classification: a complementary tool for setting indirect outcome-based analytical performance specifications
  13. Genetics and Molecular Diagnostics
  14. Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations
  15. Candidate Reference Measurement Procedures and Materials
  16. Antiphospholipid IgG Certified Reference Material ERM®-DA477/IFCC: a tool for aPL harmonization?
  17. General Clinical Chemistry and Laboratory Medicine
  18. External quality assessment of the manual tilt tube technique for prothrombin time testing: a report from the IFCC-SSC/ISTH Working Group on the Standardization of PT/INR
  19. Simple steps to achieve harmonisation and standardisation of dried blood spot phenylalanine measurements and facilitate consistent management of patients with phenylketonuria
  20. Inclusion of pyridoxine dependent epilepsy in expanded newborn screening programs by tandem mass spectrometry: set up of first and second tier tests
  21. Analytical performance evaluation and optimization of serum 25(OH)D LC-MS/MS measurement
  22. Towards routine high-throughput analysis of fecal bile acids: validation of an enzymatic cycling method for the quantification of total bile acids in human stool samples on fully automated clinical chemistry analyzers
  23. Analytical and clinical evaluations of Snibe Maglumi® S100B assay
  24. Prevalence and detection of citrate contamination in clinical laboratory
  25. Reference Values and Biological Variations
  26. Temporal dynamics in laboratory medicine: cosinor analysis and real-world data (RWD) approaches to population chronobiology
  27. Establishing sex- and age-related reference intervals of serum glial fibrillary acid protein measured by the fully automated lumipulse system
  28. Hematology and Coagulation
  29. Performance of the automated digital cell image analyzer UIMD PBIA in white blood cell classification: a comparative study with sysmex DI-60
  30. Cancer Diagnostics
  31. Flow-cytometric MRD detection in pediatric T-ALL: a multicenter AIEOP-BFM consensus-based guided standardized approach
  32. Impact of biological and genetic features of leukemic cells on the occurrence of “shark fins” in the WPC channel scattergrams of the Sysmex XN hematology analyzers in patients with chronic lymphocytic leukemia
  33. Assessing the clinical applicability of dimensionality reduction algorithms in flow cytometry for hematologic malignancies
  34. Cardiovascular Diseases
  35. Evaluation of sex-specific 0-h high-sensitivity cardiac troponin T thresholds for the risk stratification of non-ST-segment elevation myocardial infarction
  36. Retraction
  37. The first case of Teclistamab interference with serum electrophoresis and immunofixation
  38. Letters to the Editor
  39. Is this quantitative test fit-for-purpose?
  40. Reply to “Is this quantitative test fit-for-purpose?”
  41. Short-term biological variation of coagulation and fibrinolytic measurands
  42. The first case of Teclistamab interference with serum electrophoresis and immunofixation
  43. Imlifidase: a new interferent on serum protein electrophoresis looking as a rare plasma cell dyscrasia
  44. Research on the development of image-based Deep Learning (DL) model for serum quality recognition
  45. Interference of hypertriglyceridemia on total cholesterol assay with the new CHOL2 Abbott method on Architect analyser
  46. Congress Abstracts
  47. 10th Annual Meeting of the Austrian Society for Laboratory Medicine and Clinical Chemistry (ÖGLMKC)
https://doi.org/10.1515/cclm-2024-0962
Keywords for this article
ARMS-PCR; base substitution; point mutation; polymerase extension; SNP; thermodynamic stability

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. The journey to pre-analytical quality
  4. Manual tilt tube method for prothrombin time: a commentary on contemporary relevance
  5. Reviews
  6. From errors to excellence: the pre-analytical journey to improved quality in diagnostics. A scoping review
  7. Advancements and challenges in high-sensitivity cardiac troponin assays: diagnostic, pathophysiological, and clinical perspectives
  8. Opinion Paper
  9. Is it feasible for European laboratories to use SI units in reporting results?
  10. Perspectives
  11. What does cancer screening have to do with tomato growing?
  12. Computer simulation approaches to evaluate the interaction between analytical performance characteristics and clinical (mis)classification: a complementary tool for setting indirect outcome-based analytical performance specifications
  13. Genetics and Molecular Diagnostics
  14. Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations
  15. Candidate Reference Measurement Procedures and Materials
  16. Antiphospholipid IgG Certified Reference Material ERM®-DA477/IFCC: a tool for aPL harmonization?
  17. General Clinical Chemistry and Laboratory Medicine
  18. External quality assessment of the manual tilt tube technique for prothrombin time testing: a report from the IFCC-SSC/ISTH Working Group on the Standardization of PT/INR
  19. Simple steps to achieve harmonisation and standardisation of dried blood spot phenylalanine measurements and facilitate consistent management of patients with phenylketonuria
  20. Inclusion of pyridoxine dependent epilepsy in expanded newborn screening programs by tandem mass spectrometry: set up of first and second tier tests
  21. Analytical performance evaluation and optimization of serum 25(OH)D LC-MS/MS measurement
  22. Towards routine high-throughput analysis of fecal bile acids: validation of an enzymatic cycling method for the quantification of total bile acids in human stool samples on fully automated clinical chemistry analyzers
  23. Analytical and clinical evaluations of Snibe Maglumi® S100B assay
  24. Prevalence and detection of citrate contamination in clinical laboratory
  25. Reference Values and Biological Variations
  26. Temporal dynamics in laboratory medicine: cosinor analysis and real-world data (RWD) approaches to population chronobiology
  27. Establishing sex- and age-related reference intervals of serum glial fibrillary acid protein measured by the fully automated lumipulse system
  28. Hematology and Coagulation
  29. Performance of the automated digital cell image analyzer UIMD PBIA in white blood cell classification: a comparative study with sysmex DI-60
  30. Cancer Diagnostics
  31. Flow-cytometric MRD detection in pediatric T-ALL: a multicenter AIEOP-BFM consensus-based guided standardized approach
  32. Impact of biological and genetic features of leukemic cells on the occurrence of “shark fins” in the WPC channel scattergrams of the Sysmex XN hematology analyzers in patients with chronic lymphocytic leukemia
  33. Assessing the clinical applicability of dimensionality reduction algorithms in flow cytometry for hematologic malignancies
  34. Cardiovascular Diseases
  35. Evaluation of sex-specific 0-h high-sensitivity cardiac troponin T thresholds for the risk stratification of non-ST-segment elevation myocardial infarction
  36. Retraction
  37. The first case of Teclistamab interference with serum electrophoresis and immunofixation
  38. Letters to the Editor
  39. Is this quantitative test fit-for-purpose?
  40. Reply to “Is this quantitative test fit-for-purpose?”
  41. Short-term biological variation of coagulation and fibrinolytic measurands
  42. The first case of Teclistamab interference with serum electrophoresis and immunofixation
  43. Imlifidase: a new interferent on serum protein electrophoresis looking as a rare plasma cell dyscrasia
  44. Research on the development of image-based Deep Learning (DL) model for serum quality recognition
  45. Interference of hypertriglyceridemia on total cholesterol assay with the new CHOL2 Abbott method on Architect analyser
  46. Congress Abstracts
  47. 10th Annual Meeting of the Austrian Society for Laboratory Medicine and Clinical Chemistry (ÖGLMKC)
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 21.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2024-0962/html
Scroll to top button