Evidence map›Paper›PMID 28381232›Full record

ArticleJournal of translational medicine2017

Detection of newly produced T and B lymphocytes by digital PCR in blood stored dry on nylon flocked swabs.

Marion Vaglio Tessitore, Alessandra Sottini, Aldo M Roccaro, Claudia Ghidini, Simona Bernardi, Giovanni Martellosio, Federico Serana, Luisa Imberti

Open access · goldAbstract read
In one paragraph

Article in Journal of translational medicine, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.

0numbers the graph read from it
0cells of the map it votes in
7citing papers in PubMed
1.0field-weighted citation impact, top 23% of its field
1 · What the graph read from it

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.

2 · The registry

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.

3 · Its place in the literature

Who cites it

7 citing papers in PubMed, 15 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 1 institution in 1 country.

Marion Vaglio TessitoreCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Alessandra SottiniCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Aldo M RoccaroCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Claudia GhidiniCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Simona BernardiCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Giovanni MartellosioCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Federico SeranaCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy.
Luisa ImbertiCentro di Ricerca Emato-oncologica AIL (CREA), ASST Spedali Civili, P.le Spedali Civili, 1, 25123, Brescia, Italy. luisa.imberti@asst-spedalicivili.it.
Azienda Socio Sanitaria Territoriale degli Spedali Civili di Brescia · IT

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundA normal number of T-cell receptor excision circles (TRECs) and K-deleting recombination excision circles (KRECs) is considered a biomarker for adequate new T- and B-cell production. In newborns, detection of TRECs and KRECs by real time PCR from dried blood spotted on filter paper is used for the screening of severe immunodeficiency. In adults, elderly and during diseases, where the number of TRECs is lower than in newborns and children, a large amount of DNA and a sensitive method of amplification are necessary to identify newly produced lymphocytes.

methodsDNA was prepared from blood of 203 healthy adults (range: 18-91 years old) absorbed for 10 s on flocked swabs and let to dry, or from peripheral blood mononuclear cells. DNA was subjected to digital PCR and to well established conventional real time PCR-based method using TREC- and KREC-specific primers and probes. The number of TRECs and KRECs was expressed per mL of blood. Statistical analysis was performed by nested ANOVA, Pearson coefficient of determination, and by linear regression tests.

resultsThe novel method for the storage of dried blood on nylon flocked swabs and the use of digital PCR allow quantification of TRECs and KRECs with high degree of sensitivity, specificity, accuracy, and precision. TRECs and KRECs were amplified by digital PCR in all tested blood samples, including those obtained from elderly individuals (>70 years old) and that were negative by real time PCR. Furthermore, values of TRECs and KRECs obtained by digital PCR were in the range of those acquired by real time PCR.

conclusionsOur findings demonstrate that DNA isolation from dried blood on flocked swabs followed by digital PCR-based analysis represents a useful tool for studying new lymphocyte production in adults and elderly individuals. This suggests the potential use of the methodology when monitoring of clinical variables is limited by the number of molecules that can be amplified and detected, such as in patients with immunodeficiency or under immunosuppressive therapies.

Indexed as

AdolescentAdultAgedAged, 80 and overBlood Specimen CollectionB-LymphocytesDNAElectrophoresis, Agar GelHeLa CellsHumansMiddle AgedNylonsReal-Time Polymerase Chain ReactionRecombination, GeneticReproducibility of ResultsT-LymphocytesDNANylonsDigital PCRFlocked swabsKRECsTRECs

Identifiers

PMID28381232
PMCPMC5381048
OpenAlexW2604900646

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LicenceCC BY
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Registered trials

None linked

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.