Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection
Objective To explore the potential application of glucans binding protein (GBP) in the detection of soluble glucans from serum sampled from patients with invasive fungal infection (IFI). Methods The prokaryotic plasmid pET30a-GBP252-668 coding for 252 to 668 amino acid in GBP as an active domain of...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2022-06-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-6-883.pdf |
| _version_ | 1797361460236517376 |
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| author | ZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei |
| author_facet | ZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei |
| author_sort | ZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei |
| collection | DOAJ |
| description | Objective To explore the potential application of glucans binding protein (GBP) in the detection of soluble glucans from serum sampled from patients with invasive fungal infection (IFI). Methods The prokaryotic plasmid pET30a-GBP252-668 coding for 252 to 668 amino acid in GBP as an active domain of GBP of Limulus polyphemus was constructed. The recombinant GBP252-668 was expressed and purified.Then a double GBP252-668 sandwich method was developed to detect (1,3)-β-D-glucans in the serum of patients with IFI. Meanwhile, the results were compared with G-test. Results Recombinant GBP252-668 (molecular weight is about 46 ku) was obtained and the double GBP252-668 sandwich method was established and optimized via chessboard method, the concentrations of GBP252-668 were 8 and 16 g/L for coating and detection respectively. The serum sampled from 48 patients with IFI and from 48 healthy donors were examined.and results were analysed. The sensitivity and specificity of double GBP252-668 sandwich method were 91.67% and 89.58% respectively and the sensitivity was non-inferior to G-test, and specificity of double GBP252-668 sandwich method was even higher than that of G-test (79.17%). Conclusions The GBP252-668 sandwich method is a potential detection method for IFI diagnosis. The method is non-inferior to G-test and its specificity is even higher than G-test. It is expected to be a new commercially available kit for IFI detection. |
| first_indexed | 2024-03-08T15:54:02Z |
| format | Article |
| id | doaj.art-d163071d2e634c5bbc68db1f62510abd |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T15:54:02Z |
| publishDate | 2022-06-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-d163071d2e634c5bbc68db1f62510abd2024-01-09T02:51:51ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252022-06-0142688388910.16352/j.issn.1001-6325.2022.06.007Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infectionZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei01. Department of Clinical Laboratory, Xiangyang Central Hospital, the Affiliated Hospital of Hubei College of Arts and Sciences, Xiangyang 441021; ;2. Centre for Translational Medicine, the Second Affiliated Hospital of Wannan Medical College, Wuhu 241000, ChinaObjective To explore the potential application of glucans binding protein (GBP) in the detection of soluble glucans from serum sampled from patients with invasive fungal infection (IFI). Methods The prokaryotic plasmid pET30a-GBP252-668 coding for 252 to 668 amino acid in GBP as an active domain of GBP of Limulus polyphemus was constructed. The recombinant GBP252-668 was expressed and purified.Then a double GBP252-668 sandwich method was developed to detect (1,3)-β-D-glucans in the serum of patients with IFI. Meanwhile, the results were compared with G-test. Results Recombinant GBP252-668 (molecular weight is about 46 ku) was obtained and the double GBP252-668 sandwich method was established and optimized via chessboard method, the concentrations of GBP252-668 were 8 and 16 g/L for coating and detection respectively. The serum sampled from 48 patients with IFI and from 48 healthy donors were examined.and results were analysed. The sensitivity and specificity of double GBP252-668 sandwich method were 91.67% and 89.58% respectively and the sensitivity was non-inferior to G-test, and specificity of double GBP252-668 sandwich method was even higher than that of G-test (79.17%). Conclusions The GBP252-668 sandwich method is a potential detection method for IFI diagnosis. The method is non-inferior to G-test and its specificity is even higher than G-test. It is expected to be a new commercially available kit for IFI detection.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-6-883.pdfinvasive fungal infection(ifi)|glucans binding protein|sandwich method|clinical application |
| spellingShingle | ZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection Jichu yixue yu linchuang invasive fungal infection(ifi)|glucans binding protein|sandwich method|clinical application |
| title | Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| title_full | Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| title_fullStr | Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| title_full_unstemmed | Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| title_short | Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| title_sort | prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection |
| topic | invasive fungal infection(ifi)|glucans binding protein|sandwich method|clinical application |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-6-883.pdf |
| work_keys_str_mv | AT zhuliruhuangyoumingzhanghongxiangzhoufayoutangxiaolei prokaryoticexpressionofglucansbindingproteinandapplicationfordetectinginvasivefungalinfection |