Development and Assessment of a Quantitative Reverse Transcription-PCR Assay for Simultaneous Measurement of Four Amplicons
Background: High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex...
Saved in:
| Published in: | Clinical chemistry (Baltimore, Md.) Vol. 49; no. 9; pp. 1467 - 1475 |
|---|---|
| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Washington, DC
Am Assoc Clin Chem
01.09.2003
American Association for Clinical Chemistry Oxford University Press |
| Subjects: | |
| ISSN: | 0009-9147, 1530-8561 |
| Online Access: | Get full text |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Abstract | Background: High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system.
Methods: We developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl2, and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays.
Results: The simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log10 cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of −3.322.
Conclusions: This is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer. |
|---|---|
| AbstractList | Background: High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system.
Methods: We developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl2, and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays.
Results: The simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log10 cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of −3.322.
Conclusions: This is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer. High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system. We developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl(2), and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays. The simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log(10) cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of -3.322. This is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer. High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system. We developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl(2), and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays. The simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log(10) cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of -3.322. This is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer. High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system.BACKGROUNDHigh-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously reported a quantitative reverse transcription (QRT)-PCR assay for human radiation-responsive gene targets using a whole-blood ex vivo irradiation model, but we needed a multitarget assay on a smaller, less costly, real-time PCR detection system.We developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl(2), and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays.METHODSWe developed a quadruplex QRT-PCR assay in a 96-well, closed-plate format suitable for use with RNA extracted from whole blood. Four cDNA targets were simultaneously amplified in a sealed tube by hybridization to exonuclease probes, each conjugated to distinct fluorogenic reporters. A novel primer-limited 18S rRNA reference target was validated from serial dilutions of human total RNA. To test assay precision, we incorporated a positive-control cDNA mimic into duplex and quadruplex PCR reactions. The master mixture was supplemented with more enzyme, MgCl(2), and deoxyribonucleotides. Simultaneous detection of four targets was evaluated in comparison with respective duplex QRT-PCR assays.The simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log(10) cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of -3.322.RESULTSThe simultaneous detection of three radiation-responsive genes by quadruplex QRT-PCR was quantitative, with gene expression changes similar to those observed with optimized duplex and triplex QRT-PCR assays. The 18S rRNA and GADD45 calibration curves (threshold cycle vs log(10) cDNA) were linear and reproducible and showed optimal PCR efficiencies as indicated by slopes statistically equivalent to the theoretical value of -3.322.This is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer.CONCLUSIONSThis is the first study of a quadruplex QRT-PCR assay. Our approach has diagnostic utility in the detection of biomarkers, biological and toxicologic agents, and genes of inherited diseases and cancer. |
| Author | Grace, Marcy B Jackson, William E., III McLeland, Christopher B Smith, Jeffrey M Blakely, William F Gagliardi, Steven J |
| Author_xml | – sequence: 1 fullname: Grace, Marcy B – sequence: 2 fullname: McLeland, Christopher B – sequence: 3 fullname: Gagliardi, Steven J – sequence: 4 fullname: Smith, Jeffrey M – sequence: 5 fullname: Jackson, William E., III – sequence: 6 fullname: Blakely, William F |
| BackLink | http://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15069121$$DView record in Pascal Francis https://www.ncbi.nlm.nih.gov/pubmed/12928227$$D View this record in MEDLINE/PubMed |
| BookMark | eNpt0VFrFDEQB_AgLfZaffALSBAUfNhrks1uNo_HaVVoqdb6HLK5WZuSTbbJbo_ilzfXu3JQ-hQGfjP8M3OMDnzwgNA7Sua0FOUpl3M5p7wWr9CMViUpmqqmB2hGCJGFpFwcoeOUbnPJRVO_RkeUSdYwJmbo3xe4BxeGHvyItV_hRUqQ0mMZOqzxr0n70Y56tPeArzKOCfB11D6ZaIfRBl_8XF5t2vQD7kLEv20_uVF7CFPCF6DTFOFp3FmYIl70g7Mm-PQGHXbaJXi7e0_Qn7Ov18vvxfnltx_LxXlheFmORd3VKyO4JkyzSkAtWgGMQC6plrQVFcjWQFakok3dkoylrjTwrik5o6Y8QZ-2c4cY7iZIo-ptMuDcNqQSZUV4I5sMPzyDtzmwz9kUozyvj5R1Ru93aGp7WKkh2l7HB_W00ww-7oBORrsu78rYtHcVqSVlNLvPW2diSClCtydEbe6quFRSbe6a7ekzax5vEvwYtXUvduw-fWP_3qxtBJV67VwOTdV6vd7D_zT7sVs |
| CODEN | CLCHAU |
| CitedBy_id | crossref_primary_10_1097_01_HP_0000175913_36594_a4 crossref_primary_10_1259_bjr_20130173 crossref_primary_10_3389_fonc_2021_684531 crossref_primary_10_1016_j_yexmp_2007_12_001 crossref_primary_10_1016_j_foodcont_2012_11_044 crossref_primary_10_1016_j_genrep_2019_100416 crossref_primary_10_3109_09553002_2014_938374 crossref_primary_10_1016_j_fct_2006_08_021 crossref_primary_10_3390_radiation2010006 crossref_primary_10_1667_RR2738_1 crossref_primary_10_4103_0973_1482_160912 crossref_primary_10_1007_s00284_012_0154_z crossref_primary_10_3109_09553002_2010_519424 crossref_primary_10_1016_j_lfs_2003_11_010 crossref_primary_10_1016_j_jcv_2008_03_005 crossref_primary_10_1016_j_radmeas_2007_05_011 crossref_primary_10_1016_j_intimp_2009_08_012 crossref_primary_10_1089_fpd_2016_2143 crossref_primary_10_1093_jrr_rrs060 crossref_primary_10_1667_21_00064_1 crossref_primary_10_1016_j_gene_2003_08_007 crossref_primary_10_1002_cam4_132 crossref_primary_10_1088_1361_6498_ac15df crossref_primary_10_3109_09553002_2011_568574 crossref_primary_10_1097_HP_0b013e31829cf221 crossref_primary_10_1016_j_radmeas_2007_05_035 crossref_primary_10_1158_0008_5472_CAN_04_1883 crossref_primary_10_1002_elps_200900074 crossref_primary_10_1016_j_ab_2005_06_026 crossref_primary_10_1155_2011_910369 crossref_primary_10_3390_cells10113251 crossref_primary_10_1016_j_jchromb_2014_06_004 crossref_primary_10_2353_jmoldx_2006_050135 crossref_primary_10_1097_01_cji_0000154251_20125_2e crossref_primary_10_1007_s13277_009_0005_0 crossref_primary_10_3390_ijerph8124521 crossref_primary_10_1002_elps_201200372 crossref_primary_10_1093_jrr_rru074 crossref_primary_10_1016_j_ijrobp_2008_03_043 crossref_primary_10_1111_j_1478_3231_2005_01117_x crossref_primary_10_1667_RR1382_1 crossref_primary_10_1002_elps_201000091 crossref_primary_10_1016_j_radmeas_2007_05_004 crossref_primary_10_1016_j_intimp_2009_08_020 crossref_primary_10_3892_mmr_2017_6476 |
| Cites_doi | 10.1002/jbt.8 10.1006/mcpr.1999.0284 10.2144/99272rr01 10.1093/clinchem/45.2.297 10.1093/clinchem/45.8.1148 10.1677/jme.0.0250169 10.1093/clinchem/47.3.486 10.1016/S0166-0934(01)00390-1 10.1093/clinchem/48.8.1178 10.1034/j.1399-0039.2001.580405.x 10.1667/0033-7587(2000)154[0342:IOPMBI]2.0.CO;2 10.1093/carcin/22.8.1179 10.1073/pnas.96.11.6394 10.1006/mcpr.2002.0419 10.1515/CCLM.1998.045 10.1016/S0167-4781(00)00200-1 10.1093/nar/30.6.1292 10.1016/S0166-0934(01)00260-9 10.1128/AEM.64.2.669-677.1998 10.1080/09553000210158056 10.1093/clinchem/48.8.1329 10.1093/clinchem/47.6.990 |
| ContentType | Journal Article |
| Copyright | 2004 INIST-CNRS Copyright American Association for Clinical Chemistry Sep 2003 |
| Copyright_xml | – notice: 2004 INIST-CNRS – notice: Copyright American Association for Clinical Chemistry Sep 2003 |
| DBID | AAYXX CITATION IQODW CGR CUY CVF ECM EIF NPM 3V. 4U- 7QO 7RV 7TM 7U7 7X7 7XB 88E 88I 8AO 8C1 8FD 8FE 8FG 8FI 8FJ 8FK ABJCF ABUWG AEUYN AFKRA AZQEC BENPR BGLVJ BHPHI BKSAR C1K CCPQU D1I DWQXO FR3 FYUFA GHDGH GNUQQ HCIFZ K9. KB. KB0 M0S M1P M2P NAPCQ P64 PCBAR PDBOC PHGZM PHGZT PJZUB PKEHL PPXIY PQEST PQGLB PQQKQ PQUKI PRINS Q9U RC3 S0X 7X8 |
| DOI | 10.1373/49.9.1467 |
| DatabaseName | CrossRef Pascal-Francis Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed ProQuest Central (Corporate) University Readers Biotechnology Research Abstracts Nursing & Allied Health Database Nucleic Acids Abstracts Toxicology Abstracts Health & Medical Collection ProQuest Central (purchase pre-March 2016) Medical Database (Alumni Edition) Science Database (Alumni Edition) ProQuest Pharma Collection Public Health Database Technology Research Database ProQuest SciTech Collection ProQuest Technology Collection Hospital Premium Collection Hospital Premium Collection (Alumni Edition) ProQuest Central (Alumni) (purchase pre-March 2016) Materials Science & Engineering Collection ProQuest Central (Alumni) ProQuest One Sustainability (subscription) ProQuest Central UK/Ireland ProQuest Central Essentials ProQuest Central Technology Collection Natural Science Collection Earth, Atmospheric & Aquatic Science Collection Environmental Sciences and Pollution Management ProQuest One Community College ProQuest Materials Science Collection ProQuest Central Engineering Research Database Health Research Premium Collection Health Research Premium Collection (Alumni) ProQuest Central Student ProQuest SciTech Premium Collection ProQuest Health & Medical Complete (Alumni) Materials Science Database Nursing & Allied Health Database (Alumni Edition) ProQuest Health & Medical Collection Medical Database Science Database ProQuest Nursing & Allied Health Premium Biotechnology and BioEngineering Abstracts Earth, Atmospheric & Aquatic Science Database Materials Science Collection ProQuest Central Premium ProQuest One Academic (New) ProQuest Health & Medical Research Collection ProQuest One Academic Middle East (New) ProQuest One Health & Nursing ProQuest One Academic Eastern Edition (DO NOT USE) ProQuest One Applied & Life Sciences ProQuest One Academic (retired) ProQuest One Academic UKI Edition ProQuest Central China ProQuest Central Basic Genetics Abstracts SIRS Editorial MEDLINE - Academic |
| DatabaseTitle | CrossRef MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) ProQuest Central Student ProQuest Central Essentials Nucleic Acids Abstracts SciTech Premium Collection ProQuest Central China Environmental Sciences and Pollution Management ProQuest One Applied & Life Sciences ProQuest One Sustainability Health Research Premium Collection Natural Science Collection Health & Medical Research Collection ProQuest Central (New) ProQuest Medical Library (Alumni) ProQuest Science Journals (Alumni Edition) ProQuest One Academic Eastern Edition Earth, Atmospheric & Aquatic Science Database ProQuest Hospital Collection ProQuest Technology Collection Health Research Premium Collection (Alumni) ProQuest Hospital Collection (Alumni) Biotechnology and BioEngineering Abstracts Nursing & Allied Health Premium ProQuest Health & Medical Complete ProQuest One Academic UKI Edition ProQuest Nursing & Allied Health Source (Alumni) Engineering Research Database ProQuest One Academic ProQuest One Academic (New) University Readers Technology Collection Technology Research Database ProQuest One Academic Middle East (New) SIRS Editorial Materials Science Collection ProQuest Health & Medical Complete (Alumni) ProQuest Central (Alumni Edition) ProQuest One Community College ProQuest One Health & Nursing ProQuest Pharma Collection ProQuest Central Earth, Atmospheric & Aquatic Science Collection ProQuest Health & Medical Research Collection Genetics Abstracts Biotechnology Research Abstracts Health and Medicine Complete (Alumni Edition) ProQuest Central Korea Materials Science Database ProQuest Materials Science Collection ProQuest Public Health ProQuest Central Basic Toxicology Abstracts ProQuest Science Journals ProQuest Nursing & Allied Health Source ProQuest SciTech Collection ProQuest Medical Library Materials Science & Engineering Collection ProQuest Central (Alumni) MEDLINE - Academic |
| DatabaseTitleList | CrossRef MEDLINE ProQuest Central Student MEDLINE - Academic |
| Database_xml | – sequence: 1 dbid: NPM name: PubMed url: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: BENPR name: ProQuest Central url: https://www.proquest.com/central sourceTypes: Aggregation Database |
| DeliveryMethod | fulltext_linktorsrc |
| Discipline | Medicine |
| EISSN | 1530-8561 |
| EndPage | 1475 |
| ExternalDocumentID | 423236261 12928227 15069121 10_1373_49_9_1467 www49_9_1467 |
| Genre | Research Support, U.S. Gov't, Non-P.H.S Journal Article |
| GroupedDBID | - 08R 1AW 29B 2WC 3V. 4.4 53G 55 5GY 5RE 5VS 5WD 7RV 7X7 88E 88I 8AO 8C1 8FE 8FG 8FH 8FI 8FJ 8R4 8R5 AALRV AAPBV AAPQZ AAQQT ABFLS ABJCF ABOCM ABPPZ ABSAR ABSGY ABUWG ACGOD ACIWK ACPRK ADBBV ADGZP ADIPN AELNO AELWJ AENEX AETBJ AFFNX AFKRA AFRAH AGINJ AGVJH AHMBA AIMBJ AJYGW ALMA_UNASSIGNED_HOLDINGS AZQEC BAWUL BBAFP BCRHZ BENPR BES BGLVJ BHPHI BKEYQ BKSAR BPHCQ BTFSW BVXVI C1A CS3 D1I DIK DU5 DWQXO E3Z EBS EJD ENERS EX3 F5P FHSFR FOTVD FQBLK FYUFA GJ GNUQQ GX1 HCIFZ HH5 H~9 IAO IEA IGS IH2 IHR IHW INH INIJC INR ITC J5H K-O KB. KOP KQ8 L7B LK5 LOTEE LXI M1P M2P M7R NADUK O0- OBOKY OCZFY OK1 PCBAR PDBOC PQEST PQQKQ PQUKI PRINS PROAC PSQYO Q2X R0Z RHF RHI RNS ROX RZL S0X SJN TCC TWZ U5U UCJ UNMZH V8 WH7 WOQ WOW X X7M YXANX ZA5 ZCG ZE2 ZGI --- -~X .55 .GJ 04C 0R~ 18M 6J9 AABZA AACZT AAPXW AARHZ AAUAY AAVAP AAYXX ABCQX ABDFA ABEJV ABGNP ABJNI ABMNT ABNHQ ABPQP ABPTD ABQNK ABSQV ABVGC ABWST ABXVV ABXZS ACIHN ACYHN ADGKP ADNBA ADQBN ADVEK AEAQA AEMQT AEUYN AFFHD AFFZL AFGWE AFXAL AFYAG AGQXC AGUTN AHMMS AJBYB AJEEA AJNCP ALXQX ATGXG BEYMZ C45 CCPQU CITATION F9R FECEO FLUFQ FOEOM GAUVT HMCUK JXSIZ KBUDW KSI KSN NAPCQ NOMLY NU- OAUYM OJZSN OPAEJ OVD OWPYF PHGZM PHGZT PJZUB PPXIY PQGLB RUSNO TEORI TR2 UKHRP W8F YBU YHG YSK YWH ~V8 1KJ AAPGJ AAWDT ABEFU ACFRR ACUTJ ACVCV ACZBC ADMTO AFFQV AGKRT AGMDO AGORE AHGBF AI. AJDVS ALIPV APJGH AQDSO AVNTJ BMSDO EBD EIHBH EIHJH EMOBN H13 IQODW ML- MVM OBFPC PV9 SV3 TMA VH1 YQJ CGR CUY CVF ECM EIF NPM 4U- 7QO 7TM 7U7 7XB 8FD 8FK ARBBW C1K FR3 K9. P64 PKEHL Q9U RC3 7X8 PUEGO |
| ID | FETCH-LOGICAL-c433t-6f6dc74a02a257e67b7e20e02a1a91b75e9bce6f605186b0dc79a5ae4f83421c3 |
| IEDL.DBID | M2P |
| ISICitedReferencesCount | 52 |
| ISICitedReferencesURI | http://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=Summon&SrcAuth=ProQuest&DestLinkType=CitingArticles&DestApp=WOS_CPL&KeyUT=000184896200008&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| ISSN | 0009-9147 |
| IngestDate | Sun Sep 28 00:02:50 EDT 2025 Tue Oct 07 05:17:50 EDT 2025 Tue Aug 05 11:40:08 EDT 2025 Mon Jul 21 09:14:31 EDT 2025 Tue Nov 18 21:17:26 EST 2025 Sat Nov 29 02:18:59 EST 2025 Tue Nov 10 19:49:10 EST 2020 |
| IsDoiOpenAccess | false |
| IsOpenAccess | true |
| IsPeerReviewed | true |
| IsScholarly | true |
| Issue | 9 |
| Keywords | Human RNA Blood Dose activity relation Gene Reference system Amplicon DNA Clinical biology Irradiation Genetics Technique Molecular biology Reverse transcription polymerase chain reaction Comparative study Quantitative analysis |
| Language | English |
| License | https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model CC BY 4.0 |
| LinkModel | DirectLink |
| MergedId | FETCHMERGED-LOGICAL-c433t-6f6dc74a02a257e67b7e20e02a1a91b75e9bce6f605186b0dc79a5ae4f83421c3 |
| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 |
| OpenAccessLink | https://academic.oup.com/clinchem/article-pdf/49/9/1467/32731167/clinchem1467.pdf |
| PMID | 12928227 |
| PQID | 214000036 |
| PQPubID | 47786 |
| PageCount | 9 |
| ParticipantIDs | proquest_miscellaneous_73504898 proquest_journals_214000036 pubmed_primary_12928227 pascalfrancis_primary_15069121 crossref_primary_10_1373_49_9_1467 crossref_citationtrail_10_1373_49_9_1467 highwire_smallpub1_www49_9_1467 |
| ProviderPackageCode | BTFSW RHF RHI |
| PublicationCentury | 2000 |
| PublicationDate | 2003-09-01 |
| PublicationDateYYYYMMDD | 2003-09-01 |
| PublicationDate_xml | – month: 09 year: 2003 text: 2003-09-01 day: 01 |
| PublicationDecade | 2000 |
| PublicationPlace | Washington, DC |
| PublicationPlace_xml | – name: Washington, DC – name: England – name: Washington |
| PublicationTitle | Clinical chemistry (Baltimore, Md.) |
| PublicationTitleAlternate | Clin Chem |
| PublicationYear | 2003 |
| Publisher | Am Assoc Clin Chem American Association for Clinical Chemistry Oxford University Press |
| Publisher_xml | – name: Am Assoc Clin Chem – name: American Association for Clinical Chemistry – name: Oxford University Press |
| References | (2020030122324211800_R23) 1998; 26 (2020030122324211800_R2) 2002; 100 (2020030122324211800_R6) 1999; 45 (2020030122324211800_R25) 2000; 25 (2020030122324211800_R19) 2002; 167 (2020030122324211800_R20) 2002; 78 (2020030122324211800_R13) 2002; 48 (2020030122324211800_R1) 1993; 11 (2020030122324211800_R21) 2000; 154 (2020030122324211800_R24) 2000; 1494 (2020030122324211800_R15) 1999; 45 (2020030122324211800_R10) 2001; 432 (2020030122324211800_R11) 2001; 58 (2020030122324211800_R4) 2002; 30 (2020030122324211800_R12) 2001; 47 (2020030122324211800_R5) 2002; 48 (2020030122324211800_R18) 1999; 27 (2020030122324211800_R3) 2001; 93 (2020030122324211800_R7) 2001; 22 (2020030122324211800_R14) 1998; 36 (2020030122324211800_R27) 2000; 14 (2020030122324211800_R9) 1998; 64 (2020030122324211800_R8) 2001; 15 (2020030122324211800_R17) 1999; 96 (2020030122324211800_R22) 1992; 13 (2020030122324211800_R26) 2002; 16 (2020030122324211800_R16) 2001; 47 |
| References_xml | – volume: 15 start-page: 121 year: 2001 ident: 2020030122324211800_R8 publication-title: J Biochem Mol Toxicol doi: 10.1002/jbt.8 – volume: 11 start-page: 1026 year: 1993 ident: 2020030122324211800_R1 publication-title: Biotechnology – volume: 14 start-page: 57 year: 2000 ident: 2020030122324211800_R27 publication-title: Mol Cell Probes doi: 10.1006/mcpr.1999.0284 – volume: 27 start-page: 342 year: 1999 ident: 2020030122324211800_R18 publication-title: Biotechniques doi: 10.2144/99272rr01 – volume: 26 start-page: 112 year: 1998 ident: 2020030122324211800_R23 publication-title: Biotechniques – volume: 45 start-page: 297 year: 1999 ident: 2020030122324211800_R15 publication-title: Clin Chem doi: 10.1093/clinchem/45.2.297 – volume: 45 start-page: 1148 year: 1999 ident: 2020030122324211800_R6 publication-title: Clin Chem doi: 10.1093/clinchem/45.8.1148 – volume: 25 start-page: 169 year: 2000 ident: 2020030122324211800_R25 publication-title: J Mol Endocrinol doi: 10.1677/jme.0.0250169 – volume: 47 start-page: 486 year: 2001 ident: 2020030122324211800_R16 publication-title: Clin Chem doi: 10.1093/clinchem/47.3.486 – volume: 100 start-page: 27 year: 2002 ident: 2020030122324211800_R2 publication-title: J Virol Methods doi: 10.1016/S0166-0934(01)00390-1 – volume: 48 start-page: 1178 year: 2002 ident: 2020030122324211800_R5 publication-title: Clin Chem doi: 10.1093/clinchem/48.8.1178 – volume: 58 start-page: 250 year: 2001 ident: 2020030122324211800_R11 publication-title: Tissue Antigens doi: 10.1034/j.1399-0039.2001.580405.x – volume: 154 start-page: 342 year: 2000 ident: 2020030122324211800_R21 publication-title: Radiat Res doi: 10.1667/0033-7587(2000)154[0342:IOPMBI]2.0.CO;2 – volume: 22 start-page: 1179 year: 2001 ident: 2020030122324211800_R7 publication-title: Carcinogenesis doi: 10.1093/carcin/22.8.1179 – volume: 96 start-page: 6394 year: 1999 ident: 2020030122324211800_R17 publication-title: Proc Natl Acad Sci U S A doi: 10.1073/pnas.96.11.6394 – volume: 16 start-page: 257 year: 2002 ident: 2020030122324211800_R26 publication-title: Mol Cell Probes doi: 10.1006/mcpr.2002.0419 – volume: 36 start-page: 255 year: 1998 ident: 2020030122324211800_R14 publication-title: Clin Chem Lab Med doi: 10.1515/CCLM.1998.045 – volume: 1494 start-page: 23 year: 2000 ident: 2020030122324211800_R24 publication-title: Biochim Biophys Acta doi: 10.1016/S0167-4781(00)00200-1 – volume: 30 start-page: 1292 year: 2002 ident: 2020030122324211800_R4 publication-title: Nucleic Acids Res doi: 10.1093/nar/30.6.1292 – volume: 13 start-page: 604 year: 1992 ident: 2020030122324211800_R22 publication-title: Biotechniques – volume: 93 start-page: 105 year: 2001 ident: 2020030122324211800_R3 publication-title: J Virol Methods doi: 10.1016/S0166-0934(01)00260-9 – volume: 64 start-page: 669 year: 1998 ident: 2020030122324211800_R9 publication-title: Appl Environ Microbiol doi: 10.1128/AEM.64.2.669-677.1998 – volume: 78 start-page: 1011 year: 2002 ident: 2020030122324211800_R20 publication-title: Int J Radiat Biol doi: 10.1080/09553000210158056 – volume: 432 start-page: 69 year: 2001 ident: 2020030122324211800_R10 publication-title: Mutat Res – volume: 167 start-page: 20 year: 2002 ident: 2020030122324211800_R19 publication-title: Mil Med – volume: 48 start-page: 1329 year: 2002 ident: 2020030122324211800_R13 publication-title: Clin Chem doi: 10.1093/clinchem/48.8.1329 – volume: 47 start-page: 990 year: 2001 ident: 2020030122324211800_R12 publication-title: Clin Chem doi: 10.1093/clinchem/47.6.990 |
| SSID | ssj0004786 |
| Score | 1.9843205 |
| Snippet | Background: High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events.... High-throughput and forward-deployable biological dosimetry capabilities are required for tactical and medical decisions after radiologic events. We previously... |
| SourceID | proquest pubmed pascalfrancis crossref highwire |
| SourceType | Aggregation Database Index Database Enrichment Source Publisher |
| StartPage | 1467 |
| SubjectTerms | bcl-2-Associated X Protein Biological and medical sciences Biomarkers Blood DNA Repair DNA, Complementary - chemistry DNA-Binding Proteins - blood DNA-Binding Proteins - genetics Dosimetry GADD45 Proteins Gene expression Humans Intracellular Signaling Peptides and Proteins Investigative techniques, diagnostic techniques (general aspects) Irradiation Laboratories Male Medical sciences Miscellaneous. Technology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Proteins - chemistry Proteins - genetics Proteins - metabolism Proto-Oncogene Proteins - blood Proto-Oncogene Proteins - genetics Proto-Oncogene Proteins c-bcl-2 Radiation Reproducibility of Results Reverse Transcriptase Polymerase Chain Reaction - methods RNA - chemistry RNA, Ribosomal, 18S - blood RNA, Ribosomal, 18S - chemistry Superoxide Dismutase - blood Superoxide Dismutase - genetics |
| Title | Development and Assessment of a Quantitative Reverse Transcription-PCR Assay for Simultaneous Measurement of Four Amplicons |
| URI | http://www.clinchem.org/cgi/content/abstract/49/9/1467 https://www.ncbi.nlm.nih.gov/pubmed/12928227 https://www.proquest.com/docview/214000036 https://www.proquest.com/docview/73504898 |
| Volume | 49 |
| WOSCitedRecordID | wos000184896200008&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| hasFullText | 1 |
| inHoldings | 1 |
| isFullTextHit | |
| isPrint | |
| journalDatabaseRights | – providerCode: PRVPQU databaseName: Earth, Atmospheric & Aquatic Science Database customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: PCBAR dateStart: 20021201 isFulltext: true titleUrlDefault: https://search.proquest.com/eaasdb providerName: ProQuest – providerCode: PRVPQU databaseName: Health & Medical Collection customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: 7X7 dateStart: 20021201 isFulltext: true titleUrlDefault: https://search.proquest.com/healthcomplete providerName: ProQuest – providerCode: PRVPQU databaseName: Materials Science Database customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: KB. dateStart: 20021201 isFulltext: true titleUrlDefault: http://search.proquest.com/materialsscijournals providerName: ProQuest – providerCode: PRVPQU databaseName: Nursing & Allied Health Database customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: 7RV dateStart: 20021201 isFulltext: true titleUrlDefault: https://search.proquest.com/nahs providerName: ProQuest – providerCode: PRVPQU databaseName: ProQuest Central customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: BENPR dateStart: 20021201 isFulltext: true titleUrlDefault: https://www.proquest.com/central providerName: ProQuest – providerCode: PRVPQU databaseName: Public Health Database customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: 8C1 dateStart: 20021201 isFulltext: true titleUrlDefault: https://search.proquest.com/publichealth providerName: ProQuest – providerCode: PRVPQU databaseName: Science Database customDbUrl: eissn: 1530-8561 dateEnd: 20220430 omitProxy: false ssIdentifier: ssj0004786 issn: 0009-9147 databaseCode: M2P dateStart: 20021201 isFulltext: true titleUrlDefault: https://search.proquest.com/sciencejournals providerName: ProQuest |
| link | http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwpV3da9swED_Wdoy9dN9b2i0TYw97cWt92JKexhpaBqMhhG3kzciWDIXUaeNkMPrP9yTLSQvbXvYiMD7Ltu5DJ91PdwAfrVJO4jyYZJmRiVC5TlSJ6m6N5qZWPueXDcUm5HisZjM9idicNsIqe5sYDLVdVH6P_JjhSiBkT_l8dZ34olE-uBoraOzAHjo21CO6ztlkeyxShkKP3o1AnRYyJhbikh8LfaSDkbg_HfUpgj1C0rQ4SHVX3eLv7meYhs6e_OcPPIX96H-SL53APIMHrnkOj85jhP0F3NxBERHTWGI2qTvJoiaGXK9NEw6moZkkPv_TsnVk5We83v4kk9HUP2Z-E_SISXvhQYumcYt1Sy63W5K-uxo_h5gAakfpfwk_zk6_j74msUBDUgnOV0le57aSwqTMoOa7XJbSsdThJTWaljJzuqwcUqHmq7xMkVibzDhRKy4Yrfgr2G0WjXsDhLo8V5nVgtVW5KktS3QeLKsyxax0hg_gU8-noorZy30RjXkRQnKSF0IX2i9k5AA-bEivupQdfyJ63zO7aC_NfI5MpQUy7w7F8J4MbPvK0lxTRgdw2DO6iIrfFhsu4ws2d1FjfRimG-lC8gzNplYDeN1J0rZnpj2qVx78s-dDeBwQhQHo9hZ2V8u1ewcPq1-ri3Y5hB05_enbmQytwlaN6BD2Tk7HkylefTs5GgZFuQUrYhaT |
| linkProvider | ProQuest |
| linkToHtml | http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMw1V1Lb9QwEB6VgoAL78dSaC0EEpfQxHb8OCCEClWrtqsVKlJvrhM7UqVttt3sgip-E_-RsZPsthJw64HjKpPJJvk8M44_fwPwxinlJebBJM-tTLgSOlEFDndnNbOVCppfLjabkMOhOjrSoxX41e-FCbTKPibGQO0mZfhGvklxJhDVUz6enSehaVRYXO07aLSo2PMXP3DG1nzY_Yyv9y2l218Ot3aSrqlAUnLGZomohCsltym1iFYvZCE9TT3-zKzOCpl7XZQerRCtShQpGmubW88rxTjNSoZ-b8BNHoTFAlOQjpbbMGVsLBnKFowhXHZCRkyyTa7f6xiUrqa_XpI4MDJtgy-lartp_L3cjWlv-_5_9sAewL2uviaf2gHxEFZ8_QhuH3QMgsfw8xJLitjaEbuQJiWTilhyPrd13HiHaYAEfatp48ksZPQ-viajra_hNHtBsOInzUkgZdraT-YNOV1-cg3uKvw7xEbSPo7uJ_DtWm79KazWk9o_B5J5IVTuNKeV4yJ1RYHFkaNlrqiT3rIBvOtxYcpOnT00CRmbuOQomeHa6DBRkwN4vTA9ayVJ_mS00YPLNKd2PEYQZQbBcsli_Qrmlr7yVOiMZgNY64FlusDWmAWq8AKLoxiRwjJT-6SNZDmmBa0G8KxF7tIz1YG1LF_80_MG3Nk5PNg3-7vDvTW4G9mTkdT3ElZn07l_BbfK77OTZroehx-B4-uG72_kVW2K |
| linkToPdf | http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMw1V3fb9MwED6NDk28jN9QBpuFQOIlNLGd2H5ACLZVTIOqmkDam3FiR5rUpVvTDk38Zfx3nJ2k3STgbQ88RrlcEvvz3dn-fAfwykrpBPrBKE2NiLjMVCRzHO7WKGZK6XN-2VBsQoxG8vhYjdfgV3cWxtMqO5sYDLWdFn6NfEBxJhCypwzKlhUx3hu-PzuPfAEpv9HaVdNoEHLoLn_g7K1-d7CHXf2a0uH-191PUVtgICo4Y_MoKzNbCG5iahC5LhO5cDR2eJkYleQidSovHEohcmWWxyisTGocLyXjNCkY6r0F6wJjDN6D9Y_7o_HR6lCmCGUmfRCDFoWLNq0RE2zA1VsVTNR1Z9glKPb8TFNjF5VNbY2_B7_BCQ7v_sfNdw8228ibfGiGyn1Yc9UD2PjScgsews8r_CliKkvMMmkpmZbEkPOFqcKRPHQQxGe-mtWOzL2v7yxvNN498o-ZS4JzAVKfeLqmqdx0UZPT1WKsV1fi5xAT6Pw47h_Btxv59cfQq6aVewokcVkmU6s4LS3PYpvnGDZZWqSSWuEM68ObDiO6aPO2-_IhEx02IwXTXGnlp3CiDy-XomdNspI_Ce10QNP1qZlMEFCJRuBckdi-hr-VrjTOVEKTPmx1INOtyav1EmH4guVdtFV-A6ppaS1Yig5DyT48aVC80kyV5zOLZ__UvAMbiFr9-WB0uAV3Aq0ysP2eQ28-W7gXcLu4mJ_Us-12LBL4ftP4_Q2s6Hek |
| openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Development+and+assessment+of+a+quantitative+reverse+transcription-PCR+assay+for+simultaneous+measurement+of+four+amplicons&rft.jtitle=Clinical+chemistry+%28Baltimore%2C+Md.%29&rft.au=Grace%2C+Marcy+B&rft.au=McLeland%2C+Christopher+B&rft.au=Gagliardi%2C+Steven+J&rft.au=Smith%2C+Jeffrey+M&rft.date=2003-09-01&rft.pub=Oxford+University+Press&rft.issn=0009-9147&rft.eissn=1530-8561&rft.volume=49&rft.issue=9&rft.spage=1467&rft_id=info:doi/10.1373%2F49.9.1467&rft.externalDBID=HAS_PDF_LINK&rft.externalDocID=423236261 |
| thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0009-9147&client=summon |
| thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0009-9147&client=summon |
| thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0009-9147&client=summon |