Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis.
Immunological detection of pyrazine-2-carboxylic acid for the detection of pyrazinamide resistance in Mycobacterium tuberculosis.
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DOI：
10.1371/journal.pone.0241600
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Sheen P
中科院分区：
文献类型：
--
作者：
Florentini EA;Angulo N;Gilman RH;Alcántara R;Roncal E;Antiparra R;Toscano E;Vallejos K;Kirwan D;Zimic M;Sheen P
结核分枝杆菌（Mtb）中的吡嗪酰胺（PZA）敏感性测试是当前开发的领域，并且PZA耐药菌株越来越普遍。以前的研究已经证明，吡嗪酸（POA），由PZA脱酰胺产生的代谢产物的检测，是一个很好的预测PZA耐药，因为耐药菌株不会转化为POA在一个关键的需要的速度，而敏感菌株会做，驱逐POA到细胞外环境在一定的速度，并允许定量这种积累的分析物。为了定量POA，开发了使用针对POA的超免疫多克隆兔血清的间接竞争ELISA（icELISA）测试：为此目的，首先将纯POA共价连接到高免疫原性的匙孔帽贝Hempet，并接种在兔中。将由连接至纯POA并固定在威尔斯孔底部的牛血清白蛋白（BSA）制成的构建体用作针对加标样品和液体Mtb培养物上清液的竞争物。当分析加标样品（单独的商业POA）时，半数最大抑制浓度（IC 50）为1.16 mg/mL，检测限为200 μg/mL，并且该测定具有特异性（未检测到PZA，IC 50> 20 mg/mL）。然而，培养上清液（7 H9-OADC-PANTA培养基）破坏了竞争，并且不能获得适当的icELISA曲线。我们认为，虽然我们已经表明，它是可行的，以诱导抗POA抗体，基质效应可能会损害其分析的有用性，多个，即将到来的方法来解决这个障碍的建议。
Pyrazinamide (PZA) susceptibility testing in Mycobacterium tuberculosis (Mtb) is a current area of development and PZA-resistant strains are increasingly prevalent. Previous studies have demonstrated that the detection of pyrazinoic acid (POA), the metabolite produced by the deamidation of PZA, is a good predictor for PZA resistance since a resistant strain would not convert PZA into POA at a critical required rate, whereas a susceptible strain will do, expelling POA to the extracellular environment at a certain rate, and allowing for quantification of this accumulated analyte. In order to quantify POA, an indirect competitive ELISA (icELISA) test using hyperimmune polyclonal rabbit serum against POA was developed: for this purpose, pure POA was first covalently linked to the highly immunogenic Keyhole Limpet Hemocyanine, and inoculated in rabbits. A construct made of bovine serum albumin (BSA) linked to pure POA and fixed at the bottom of wells was used as a competitor against spiked samples and liquid Mtb culture supernatants. When spiked samples (commercial POA alone) were analyzed, the half maximal inhibitory concentration (IC50) was 1.16 mg/mL, the limit of detection 200 μg/mL and the assay was specific (it did not detect PZA, IC50 > 20 mg/mL). However, culture supernatants (7H9-OADC-PANTA medium) disrupted the competition and a proper icELISA curve was not obtainable. We consider that, although we have shown that it is feasible to induce antibodies against POA, matrix effects could damage its analytical usefulness; multiple, upcoming ways to solve this obstacle are suggested.
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影响因子：
5
作者：
BELANGER, L;SYLVESTRE, C;DUFOUR, D
通讯作者：
DUFOUR, D
影响因子：
9.4
作者：
Demers, Anne-Marie;Venter, Amour;Diacon, Andreas H.
通讯作者：
Diacon, Andreas H.
荧光生物降解聚合物生物材料的设计策略。
DOI：
10.1039/c2tb00071g
发表时间：
2013-01-14
期刊：
Journal of materials chemistry. B
影响因子：
--
作者：
Zhang Y;Yang J
通讯作者：
Yang J
影响因子：
3.2
作者：
Meinzen, Christopher;Proano, Alvaro;Sheen, Patricia
通讯作者：
Sheen, Patricia
影响因子：
3
作者：
Zhang, Y;Permar, S;Sun, ZH
通讯作者：
Sun, ZH