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Pregled bibliografske jedinice broj: 1023747

Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma


Jurasović, Jasna; Pašalić, Daria; Sekovanić, Ankica; Orct, Tatjana; Dorotić, Adrijana; Stasenko, Sandra; Mioč, Tatjana; Miškulin, Iva; Škrgatić, Lana; Piasek, Martina
Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma // Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB2019 "Crossroads in Life Sciences", HDBMB2019 / Katalinić, Maja ; Dulić, Morana ; Stuparević, Igor (ur.).
Zagreb, 2019. str. 92-92 (poster, domaća recenzija, sažetak, znanstveni)


CROSBI ID: 1023747 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma

Autori
Jurasović, Jasna ; Pašalić, Daria ; Sekovanić, Ankica ; Orct, Tatjana ; Dorotić, Adrijana ; Stasenko, Sandra ; Mioč, Tatjana ; Miškulin, Iva ; Škrgatić, Lana ; Piasek, Martina

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB2019 "Crossroads in Life Sciences", HDBMB2019 / Katalinić, Maja ; Dulić, Morana ; Stuparević, Igor - Zagreb, 2019, 92-92

ISBN
978-953-95551-7-5

Skup
Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB2019 "Crossroads in Life Sciences", HDBMB2019

Mjesto i datum
Lovran, Hrvatska, 25-28.09.2019

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
candidate micro RNAs ; maternal blood plasma ; cord blood plasma ; quantitative RT-PCR ; tobacco smoke exposure

Sažetak
Circulating extracellular microRNAs (miRNAs) in the serum or plasma has generated great interest as a promising biomarker for diverse pathological conditions - changes in miRNA expression in body fluids develop earlier than conventional biomarkers. Their expression in mother-newborn pairs may signal uterine exposure to environmental pollutants, including tobacco smoke (TS) and toxic metals, and the role of miRNAs in fetal programming remains largely unstudied. Assessment of extracellular miRNAs in plasma is limited and remains challenging due to their low amounts in circulation, small size and the lack of reference miRNA. Preamplification of the miRNA’s cDNA can be used to facilitate reliable quantification. Within our research, we assess daily exposure to main toxic metals and maternal susceptibility as factors of developmental origins of health and disease. Preliminary results on candidate miRNAs indicated aberrant expression of miR-21 and miR- 146a in maternal plasma related to TS exposure, whereas miR-1537 and miR-190b were not expressed or had a threshold cycle (Ct) value >35 in 95% and 50% of analyzed samples. We further optimized the protocol for circulating miRNA analysis (by Qiagen miRNA assays) and tested the preamplification step to increase the sensitivity of qPCR analysis and the number of detectable miRNAs in 19 paired maternal and umbilical cord blood plasma samples. By using the ∆∆Ct method of relative quantification, we analyzed fold change for miRNA expression in the samples from healthy smoking (n=10) and non-smoking (n=9) postpartum women collected immediately after delivery. Preamplification facilitated the detection of all assayed candidate miRNAs and yielded a mean Ct improvement of 6.7±1.1 (p<0.01) compared with Ct values derived from the direct qPCR method. Values of ∆Ct between these methods were highly correlated (r>0.9) for miR-16, miR-21 and miR- 146a (whose Ct values by direct qPCR were already <35). Moreover, fold changes for miR-1537 and miR-190b in cord blood plasma were 3.5 and 1.6 indicating their upregulation related to TS exposure. Our results confirm that preamplification is a useful and reliable procedure to facilitate extracellular miRNA expression detection in maternal and cord blood plasma.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti, Kliničke medicinske znanosti



POVEZANOST RADA


Projekti:
HRZZ-IP-2016-06-1998 - Procjena svakodnevne izloženosti metalima i osobne osjetljivosti majke kao čimbenika razvojnoga podrijetla zdravlja i bolesti (METALORIGINS) (Piasek, Martina; Jurasović, Jasna, HRZZ - 2016-06) ( POIROT)

Ustanove:
Institut za medicinska istraživanja i medicinu rada, Zagreb,
Klinička bolnica "Merkur",
Medicinski fakultet, Zagreb,
Klinička bolnica "Sveti Duh",
Klinički bolnički centar Zagreb


Citiraj ovu publikaciju:

Jurasović, Jasna; Pašalić, Daria; Sekovanić, Ankica; Orct, Tatjana; Dorotić, Adrijana; Stasenko, Sandra; Mioč, Tatjana; Miškulin, Iva; Škrgatić, Lana; Piasek, Martina
Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma // Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB2019 "Crossroads in Life Sciences", HDBMB2019 / Katalinić, Maja ; Dulić, Morana ; Stuparević, Igor (ur.).
Zagreb, 2019. str. 92-92 (poster, domaća recenzija, sažetak, znanstveni)
Jurasović, J., Pašalić, D., Sekovanić, A., Orct, T., Dorotić, A., Stasenko, S., Mioč, T., Miškulin, I., Škrgatić, L. & Piasek, M. (2019) Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma. U: Katalinić, M., Dulić, M. & Stuparević, I. (ur.)Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB2019 "Crossroads in Life Sciences", HDBMB2019.
@article{article, author = {Jurasovi\'{c}, Jasna and Pa\v{s}ali\'{c}, Daria and Sekovani\'{c}, Ankica and Orct, Tatjana and Doroti\'{c}, Adrijana and Stasenko, Sandra and Mio\v{c}, Tatjana and Mi\v{s}kulin, Iva and \v{S}krgati\'{c}, Lana and Piasek, Martina}, year = {2019}, pages = {92-92}, keywords = {candidate micro RNAs, maternal blood plasma, cord blood plasma, quantitative RT-PCR, tobacco smoke exposure}, isbn = {978-953-95551-7-5}, title = {Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma}, keyword = {candidate micro RNAs, maternal blood plasma, cord blood plasma, quantitative RT-PCR, tobacco smoke exposure}, publisherplace = {Lovran, Hrvatska} }
@article{article, author = {Jurasovi\'{c}, Jasna and Pa\v{s}ali\'{c}, Daria and Sekovani\'{c}, Ankica and Orct, Tatjana and Doroti\'{c}, Adrijana and Stasenko, Sandra and Mio\v{c}, Tatjana and Mi\v{s}kulin, Iva and \v{S}krgati\'{c}, Lana and Piasek, Martina}, year = {2019}, pages = {92-92}, keywords = {candidate micro RNAs, maternal blood plasma, cord blood plasma, quantitative RT-PCR, tobacco smoke exposure}, isbn = {978-953-95551-7-5}, title = {Study of preamplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs in maternal and cord blood plasma}, keyword = {candidate micro RNAs, maternal blood plasma, cord blood plasma, quantitative RT-PCR, tobacco smoke exposure}, publisherplace = {Lovran, Hrvatska} }




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