ArticleBMC research notes2011
False negative results from using common PCR reagents.
Article in BMC research notes, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
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.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
18 citing papers in PubMed.
- Development of a Direct Cell-to-PCR Lysis Buffer Using Optimized Non-Ionic Detergents for RNA-Extraction-Free RT-qPCR.Methods and protocols · 2026Article
- Living in each other's pockets: insights into the life cycle ofIMA fungus · 2026Article
- The evolution of immune profiling: will there be a role for nanoparticles?Nanoscale horizons · 2024Review
- A Diagnostic Algorithm for Detection of Leishmania spp. in Human Fresh and Fixed Tissue Samples.The American journal of tropical medicine and hygiene · 2024Article
- Current and Future Advances in the Detection and Surveillance of Biosecurity-Relevant Equine Bacterial Diseases Using Loop-Mediated Isothermal Amplification (LAMP).Animals : an open access journal from MDPI · 2023Review
- Development of Cooperative Primer-Based Real-Time PCR Assays for the Detection of Plasmodium malariae and Plasmodium ovale.The Journal of molecular diagnostics : JMD · 2021Article
- Improved triplex real-time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat.Food science & nutrition · 2021Article
- A Novel PCR Method for Detecting ACE Gene Insertion/Deletion Polymorphisms and its Clinical Application.Biological procedures online · 2021Article
- Real-Time PCR Assay for the Diagnosis and Quantification of Co-infections byFrontiers in plant science · 2021Article
- A retrospective study of the initial 25 COVID-19 patients in Luoyang, China.Japanese journal of radiology · 2020Article
- Rapid Detection of IgM Antibodies against the SARS-CoV-2 Virus via Colloidal Gold Nanoparticle-Based Lateral-Flow Assay.ACS omega · 2020Article
- A Review of Corneal Endotheliitis and Endotheliopathy: Differential Diagnosis, Evaluation, and Treatment.Ophthalmology and therapy · 2019Review
- A Clinical TB Detection Method Based on Molecular Typing Technique with Quality Control.Computational and mathematical methods in medicine · 2019Article
- Diagnosis of Human Immunodeficiency Virus Infection.Clinical microbiology reviews · 2019Review
- Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay.Journal of clinical microbiology · 2017Article
- The saga of XMRV: a virus that infects human cells but is not a human virus.Emerging microbes & infections · 2014Review
- Absence of XMRV and closely related viruses in primary prostate cancer tissues used to derive the XMRV-infected cell line 22Rv1.PloS one · 2012Article
- Sensitivity of PCR assays for murine gammaretroviruses and mouse contamination in human blood samples.PloS one · 2012Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors.
Funding
Abstract
backgroundThe sensitivity of the PCR reaction makes it ideal for use when identifying potentially novel viral infections in human disease. Unfortunately, this same sensitivity also leaves this popular technique open to potential contamination with previously amplified PCR products, or "carry-over" contamination. PCR product carry-over contamination can be prevented with uracil-DNA-glycosylase (UNG), and it is for this reason that it is commonly included in many commercial PCR master-mixes. While testing the sensitivity of PCR assays to detect murine DNA contamination in human tissue samples, we inadvertently discovered that the use of this common PCR reagent may lead to the production of false-negative PCR results.
findingsWe show here that contamination with minute quantities of UNG-digested PCR product or any negative control PCR reactions containing primer-dimers regardless of UNG presence can completely block amplification from as much as 60 ng of legitimate target DNA.
conclusionsThese findings could potentially explain discrepant results from laboratories attempting to amplify MLV-related viruses including XMRV from human samples, as none of the published reports used internal-tube controls for amplification. The potential for false negative results needs to be considered and carefully controlled in PCR experiments, especially when the target copy number may be low - just as the potential for false positive results already is.
Identifiers
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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.