Eka Noneng Nawangsih, Lia Siti Halimah, Euis Reni Yuslianti, Sayu Putu Yuni Paryati
{"title":"开发一种新型候选诊断试剂盒,用于检测登革热抗体,使用IgY和金黄色葡萄球菌阳性蛋白","authors":"Eka Noneng Nawangsih, Lia Siti Halimah, Euis Reni Yuslianti, Sayu Putu Yuni Paryati","doi":"10.35990/amhs.v1n3.p112-120","DOIUrl":null,"url":null,"abstract":"Early diagnosis of dengue hemorrhagic fever (DHF) is very important in determining further \nmanagement. Diagnostic kits are intended as relatively faster, easier, and cheaper diagnostic \ntools. The objectives of this experimental laboratory study was to prepare a diagnostic kit \ncandidate to detect dengue antibody in human serum using co -agglutination methods by \nutilizing protein A positive Staphylococcus aureus as a carrier to bind Fc immunoglobulin \nfractions without altering its ability to bind antigen. To enhance the sensitivity and specificity \nof the diagnostic kit, chicken IgY was used as one of the components. Rabbit immunoglobulin \nchicken anti-IgY was used as an intermediary connection between protein A positive and \nStaphylococcus aureus with chicken IgY. Solution A contains Staphylococcus aureus Cowan \nI and rabbit immunoglobulin chicken anti-IgY serum while Solution B contains anti-dengue \nchicken serum and dengue antigen. The laboratory experiments produced a formula of A: B = \n1~3: 1~3 (v/v), which was then tested to human serum and compared with IgM and IgG ELISA \ntests as the gold standard. Data on the results of the test were statistically analyzed using chi - \nsquare test. Of the 65 qualified samples used, 47.69% resulted in true positive and 41.54% \nresulted in true negative. Meanwhile, the remaining samples demonstrated false negative \n(6.15%), positive predictive value (91.2%) and negative predictive value (87.1%). The \nsensitivity and specificity were 89% and 90%, respectively, with no significant diffe rences \n(p>0.05). Therefore, co-agglutination method using S. aureus with protein A positive as a \ncarrier can be proposed as a diagnostic kit candidate to detect human serum dengue antibody to \nscreen dengue hemorrhagic fever cases.","PeriodicalId":171840,"journal":{"name":"ACTA Medical Health Sciences","volume":"11 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2023-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a novel diagnostic kit candidate to detect dengue antibody, \\nusing IgY and protein a positive Staphylococcus aureus\",\"authors\":\"Eka Noneng Nawangsih, Lia Siti Halimah, Euis Reni Yuslianti, Sayu Putu Yuni Paryati\",\"doi\":\"10.35990/amhs.v1n3.p112-120\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Early diagnosis of dengue hemorrhagic fever (DHF) is very important in determining further \\nmanagement. Diagnostic kits are intended as relatively faster, easier, and cheaper diagnostic \\ntools. The objectives of this experimental laboratory study was to prepare a diagnostic kit \\ncandidate to detect dengue antibody in human serum using co -agglutination methods by \\nutilizing protein A positive Staphylococcus aureus as a carrier to bind Fc immunoglobulin \\nfractions without altering its ability to bind antigen. To enhance the sensitivity and specificity \\nof the diagnostic kit, chicken IgY was used as one of the components. Rabbit immunoglobulin \\nchicken anti-IgY was used as an intermediary connection between protein A positive and \\nStaphylococcus aureus with chicken IgY. Solution A contains Staphylococcus aureus Cowan \\nI and rabbit immunoglobulin chicken anti-IgY serum while Solution B contains anti-dengue \\nchicken serum and dengue antigen. The laboratory experiments produced a formula of A: B = \\n1~3: 1~3 (v/v), which was then tested to human serum and compared with IgM and IgG ELISA \\ntests as the gold standard. Data on the results of the test were statistically analyzed using chi - \\nsquare test. Of the 65 qualified samples used, 47.69% resulted in true positive and 41.54% \\nresulted in true negative. Meanwhile, the remaining samples demonstrated false negative \\n(6.15%), positive predictive value (91.2%) and negative predictive value (87.1%). The \\nsensitivity and specificity were 89% and 90%, respectively, with no significant diffe rences \\n(p>0.05). 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引用次数: 0
摘要
登革出血热(DHF)的早期诊断对于确定进一步的治疗非常重要。诊断试剂盒是一种相对更快、更容易、更便宜的诊断工具。本实验实验室研究的目的是利用蛋白a阳性金黄色葡萄球菌作为载体,在不改变其结合抗原的能力的情况下结合Fc免疫球蛋白部分,制备一种候选诊断试剂盒,用于用共凝集法检测人血清中的登革热抗体。为提高诊断试剂盒的灵敏度和特异性,采用鸡IgY作为其中一种成分。兔免疫球蛋白鸡抗IgY作为A蛋白阳性和金黄色葡萄球菌与鸡IgY之间的中介联系。溶液A含有金黄色葡萄球菌考恩I和兔免疫球蛋白鸡抗igy血清,溶液B含有抗登革热鸡血清和登革热抗原。实验室实验得到了a: B = 1~ 3:1 ~3 (v/v)的计算公式,并与IgM和IgG ELISA试验作为金标准进行了比较。检验结果的资料采用卡方检验进行统计分析。65份合格样本中,真阳性率为47.69%,真阴性为41.54%。剩余样本的预测结果为假阴性(6.15%)、阳性预测值(91.2%)和阴性预测值(87.1%)。敏感性为89%,特异性为90%,差异无统计学意义(p < 0.05)。因此,可以提出以A蛋白阳性金黄色葡萄球菌为载体的共凝集法作为检测人血清登革抗体筛查登革出血热病例的候选诊断试剂盒。
Development of a novel diagnostic kit candidate to detect dengue antibody,
using IgY and protein a positive Staphylococcus aureus
Early diagnosis of dengue hemorrhagic fever (DHF) is very important in determining further
management. Diagnostic kits are intended as relatively faster, easier, and cheaper diagnostic
tools. The objectives of this experimental laboratory study was to prepare a diagnostic kit
candidate to detect dengue antibody in human serum using co -agglutination methods by
utilizing protein A positive Staphylococcus aureus as a carrier to bind Fc immunoglobulin
fractions without altering its ability to bind antigen. To enhance the sensitivity and specificity
of the diagnostic kit, chicken IgY was used as one of the components. Rabbit immunoglobulin
chicken anti-IgY was used as an intermediary connection between protein A positive and
Staphylococcus aureus with chicken IgY. Solution A contains Staphylococcus aureus Cowan
I and rabbit immunoglobulin chicken anti-IgY serum while Solution B contains anti-dengue
chicken serum and dengue antigen. The laboratory experiments produced a formula of A: B =
1~3: 1~3 (v/v), which was then tested to human serum and compared with IgM and IgG ELISA
tests as the gold standard. Data on the results of the test were statistically analyzed using chi -
square test. Of the 65 qualified samples used, 47.69% resulted in true positive and 41.54%
resulted in true negative. Meanwhile, the remaining samples demonstrated false negative
(6.15%), positive predictive value (91.2%) and negative predictive value (87.1%). The
sensitivity and specificity were 89% and 90%, respectively, with no significant diffe rences
(p>0.05). Therefore, co-agglutination method using S. aureus with protein A positive as a
carrier can be proposed as a diagnostic kit candidate to detect human serum dengue antibody to
screen dengue hemorrhagic fever cases.