ArticleJournal of clinical microbiology2001
Purification and characterization of PCR-inhibitory components in blood cells.
Article in Journal of clinical microbiology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 334 papers, 2 of them syntheses that pooled it.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
334 citing papers in PubMed, 2 syntheses or guidelines pooled it, 937 citations in OpenAlex.
- What do we really know about brucellosis diagnosis in livestock worldwide? A systematic review.PLoS neglected tropical diseases · 2025Pooled it
- Identification of infection by Leishmania spp. in wild and domestic animals in Brazil: a systematic review with meta-analysis (2001-2021).Parasitology research · 2023Pooled it
- Circulating microRNA profiles predict the severity of COVID-19 in hospitalized patients.Translational research : the journal of laboratory and clinical medicine · 2021Trial
- Development of Universal Primer Sets for Zika Virus Envelope Gene Amplification and Sequencing.International journal of molecular sciences · 2026Article
- PVA sponge method improves detection efficiency of cfDNA in plasma.International journal of clinical oncology · 2026Article
- Unearthing forensic genetics: preliminary results on the ability to generate DNA profiles from buried human biological samples.International journal of legal medicine · 2026Article
- Substrate Recognition Governs Reverse Transcriptase Resistance to Diagnostic Inhibitors in RT-qPCR.Diagnostics (Basel, Switzerland) · 2026Article
- Non-invasive comparative study of HLA genotyping between urinary and blood DNA using sequencing-based typing and third-generation sequencing.Advances in laboratory medicine · 2026Article
- Babesiosis and Its Significance in Transfusion Medicine from a European Point of View.Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2026Review
- Analytical validation of a multiplex PCR assay for differentiation of five clinically significant nontuberculous mycobacterial species.Frontiers in microbiology · 2026Article
- Characterization of β‑lactoglobulin and κ‑casein genotypes PCR-RFLP in dairy cattle from Panama.Brazilian journal of veterinary medicine · 2026Article
- Article
- Development and diagnostic performance of a sensitive multiplex pneumonia pathogen identification assay for emergency department patients with pneumonia.Microbiology spectrum · 2025Article
- Scalable and cost-effective methods for xenomonitoring of P. falciparum and antimalarial drug resistance validated with laboratory and wild-caught mosquitoes.Scientific reports · 2025Article
- Transforming Porous Membranes into Dual-Gradient Janus Structures by Directional Asymmetric Modification.Small methods · 2025Article
- Rickettsioses as Underrecognized Cause of Hospitalization for Febrile Illness, Uganda.Emerging infectious diseases · 2025Article
- Improved HIV-1 RNA detection using whole blood versus plasma in antiretroviral-treated individuals.Journal of clinical microbiology · 2025Article
- Nanomaterials in PCR: exploring light-to-heat conversion mechanisms and microfluidic integration.Microsystems & nanoengineering · 2025Review
- Microfluidic purification of genomic DNA.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Ultrasensitive detection of intact SARS-CoV-2 particles in complex biofluids using microfluidic affinity capture.Science advances · 2025Article
274 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In a recent study, immunoglobulin G in human plasma was identified as a major inhibitor of diagnostic PCR (W. Abu Al-Soud, L. J. Jönsson, and P. Rådström. J. Clin. Microbiol. 38:345-350, 2000). In this study, two major PCR inhibitors in human blood cells were purified using size exclusion and anion-exchange chromatographic procedures. Based on N-terminal amino acid sequencing and electrophoretic analysis of the purified polypeptides, hemoglobin and lactoferrin were identified as PCR-inhibitor components in erythrocytes and leukocytes, respectively. When different concentrations of hemoglobin or lactoferrin were added to PCR mixtures of 25 microl containing 10 different thermostable DNA polymerases and 1 ng of Listeria monocytogenes DNA as template DNA, AmpliTaq Gold, Pwo, and Ultma were inhibited in the presence of < or = 1.3 microg of hemoglobin and < or = 25 ng of lactoferrin, while rTth and Tli were found to resist inhibition of at least 100 microg of hemoglobin. In addition, the quantitative effects of seven low-molecular-mass inhibitors, present in blood samples or degradation products of hemoglobin, on real-time DNA synthesis of rTth using the LightCycler Instrument were investigated. A reaction system based on a single-stranded poly(dA) template with an oligo(dT) primer annealed to the 3' end was used. It was found that the addition of 0.25 to 0.1 mg of bile per ml, 2.5 mM CaCl2, 0.25 mM EDTA, 5 microM FeCl3, and 0.01 IU of heparin per ml reduced the fluorescence to approximately 76, 70, 46, 17, and 51%, respectively. Finally, the effects of nine amplification facilitators were studied in the presence of hemoglobin and lactoferrin. Bovine serum albumin (BSA) was the most efficient amplification facilitator, so that the addition of 0.4% (wt/vol) BSA allowed AmpliTaq Gold to amplify DNA in the presence of 20 instead of 1 microg of hemoglobin and 500 instead of 5 ng of lactoferrin. Including 0.02% (wt/vol) gp32, a single-stranded-DNA binding protein, in the reaction mixture of AmpliTaq Gold was also found to reduce the inhibitory effects of hemoglobin and lactoferrin.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.