{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T06:15:38Z","timestamp":1781849738777,"version":"3.54.5"},"reference-count":56,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2022,8,30]],"date-time":"2022-08-30T00:00:00Z","timestamp":1661817600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Characteristic Innovation Project of Colleges and Universities of the Guangdong Provincial Department of Education","award":["2020KTSCX017"],"award-info":[{"award-number":["2020KTSCX017"]}]},{"name":"Characteristic Innovation Project of Colleges and Universities of the Guangdong Provincial Department of Education","award":["2022Z124"],"award-info":[{"award-number":["2022Z124"]}]},{"name":"Characteristic Innovation Project of Colleges and Universities of the Guangdong Provincial Department of Education","award":["jxsq2018102116"],"award-info":[{"award-number":["jxsq2018102116"]}]},{"name":"Ningbo Key Research and Development Plan and \u201cJie Bang Gua Shuai\u201d Program","award":["2020KTSCX017"],"award-info":[{"award-number":["2020KTSCX017"]}]},{"name":"Ningbo Key Research and Development Plan and \u201cJie Bang Gua Shuai\u201d Program","award":["2022Z124"],"award-info":[{"award-number":["2022Z124"]}]},{"name":"Ningbo Key Research and Development Plan and \u201cJie Bang Gua Shuai\u201d Program","award":["jxsq2018102116"],"award-info":[{"award-number":["jxsq2018102116"]}]},{"name":"Jiangxi Province 2018 \u2018Double Thousands Talents\u2019 Plan","award":["2020KTSCX017"],"award-info":[{"award-number":["2020KTSCX017"]}]},{"name":"Jiangxi Province 2018 \u2018Double Thousands Talents\u2019 Plan","award":["2022Z124"],"award-info":[{"award-number":["2022Z124"]}]},{"name":"Jiangxi Province 2018 \u2018Double Thousands Talents\u2019 Plan","award":["jxsq2018102116"],"award-info":[{"award-number":["jxsq2018102116"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>The tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae) is an important public-health pest that feeds on the blood of humans and some other animals. To explore the function of the target genes of C. hemipterus, it is essential to select suitable reference genes for the accurate quantification of gene expression. Here, we selected 10 frequently used reference genes in insects and evaluated their stability in C. hemipterus under various biotic (developmental stage, sex, and tissue) and abiotic (gas stimulation and temperature) conditions through RefFinder (which integrates four computational programs: geNorm, NormFinder, BestKeeper, and \u2206Ct). Results indicate that the optimal combination of reference genes for each experimental condition was as follows: RPL8 and EF1\u03b1 for the developmental stage (eggs, early instar nymphs, late instar nymphs, and adults), RPL8 and RPS16 for adult sex, RPL8 and RPL11 for adult tissue (head, thorax, abdomen, and legs), RPL8 and \u03b2-tubulin for gas stimulation (air and carbon dioxide), and RPL8 and NADH for temperature (0, 5, 17, 30, and 37 \u00b0C). Finally, the expression pattern of the HSP70 and GR21 genes were analyzed, and the results highlight the importance of appropriate reference-gene selection. Our results provide a comprehensive list of optimal reference genes from C. hemipterus for the first time, which will contribute to accurately analyzing the expression of target genes.<\/jats:p>","DOI":"10.3390\/insects13090784","type":"journal-article","created":{"date-parts":[[2022,8,31]],"date-time":"2022-08-31T00:13:56Z","timestamp":1661904836000},"page":"784","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Identification and Validation of Reference Genes for Expression Analysis Using qRT-PCR in Cimex hemipterus (Hemiptera: Cimicidae)"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8804-1373","authenticated-orcid":false,"given":"Delong","family":"Kong","sequence":"first","affiliation":[{"name":"Department of Entomology, South China Agricultural University, Guangzhou 510642, China"},{"name":"Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China"},{"name":"Engineering Research Center of Biological Control, Ministry of Education, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daxia","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Entomology, South China Agricultural University, Guangzhou 510642, China"},{"name":"Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China"},{"name":"Engineering Research Center of Biological Control, Ministry of Education, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0214-7236","authenticated-orcid":false,"given":"Changlu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Entomology, Rutgers University, New Brunswick, NJ 08901, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruyue","family":"Zhai","sequence":"additional","affiliation":[{"name":"Department of Entomology, South China Agricultural University, Guangzhou 510642, China"},{"name":"Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China"},{"name":"Engineering Research Center of Biological Control, Ministry of Education, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingling","family":"Lyu","sequence":"additional","affiliation":[{"name":"Ningbo Dayang Technology Co., Ltd., Ningbo 315000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yurong","family":"He","sequence":"additional","affiliation":[{"name":"Department of Entomology, South China Agricultural University, Guangzhou 510642, China"},{"name":"Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China"},{"name":"Engineering Research Center of Biological Control, Ministry of Education, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1979-5985","authenticated-orcid":false,"given":"Desen","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Entomology, South China Agricultural University, Guangzhou 510642, China"},{"name":"Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China"},{"name":"Engineering Research Center of Biological Control, Ministry of Education, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. 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