Testy immunologiczne enzymatyczne (EIA) do serodiagnozy ludzkiej leptospirozy: swoiste izotypowanie IgG3/IgG1 może dodatkowo pomóc w diagnostyce ostrej choroby
Pracochłonny mikroskopowy test aglutynacji (MAT) jest złotym standardem serologicznym w diagnostyce laboratoryjnej leptospirozy. Opracowaliśmy testy serologiczne oparte na EIA z wykorzystaniem białek rekombinowanych (rLigA, rLigB, rLipL32) i ekstraktów całych komórek z ośmiu serotypów Leptospira jako antygenu i oceniliśmy wydajność diagnostyczną nowego testu w każdej klasie w stosunku do paneli surowic ludzkich MAT+ (MAT+) z Portugalia/PT (n = 143) i Angola/AO (n = 100).
Stwierdziliśmy, że kombinacja rekombinowanych białek rLigA, rLigB i rLipL32 prawidłowo zidentyfikowała antygenowo swoiste IgG od pacjentów z klinicznie i laboratoryjnie potwierdzoną leptospirozą (MAT+) z 92% czułością i ~97% swoistością (AUC 0,974) w surowicy z prowincji Luanda (LDA) i Huambo (HBO) w Angoli. Połączenie ekstraktów z całych komórek L. interrogans sv Copenhageni (LiC), L. kirschneri Mozdok (LkM), L. borgpetersenii Arborea (LbA) i L. biflexa Patoc (LbP) pozwoliło dokładnie zidentyfikować pacjentów z klinicznie i laboratoryjnie potwierdzoną leptospirozą (MAT+). ) ze 100% czułością i ~98% specyficznością dla wszystkich prowincji Angola i Portugalia (AUC: 0,997 dla AO/LDA/HBO, 1,000 dla AO/HLA, 0,999 dla PT/AZ i 1,000 dla PT/LIS). Co ciekawe,
Biorąc pod uwagę, że izotypy specyficzne dla klasy IgM/IgD i IgG3/IgG1 powstają w najwcześniejszym przebiegu zakażenia, izotypowanie immunoglobuliny G może być wykorzystane do ustalenia rozpoznania ostrej leptospirozy.
- Szybkość, łatwość użycia i dokładność testów EIA sprawiają, że są one doskonałą alternatywą dla pracochłonnej i drogiej MAT do badań przesiewowych ostrej infekcji w obszarach, w których krążące serotypy patogennej Leptospira są dobrze zdefiniowane.
- W sierpniu 2020 r. nowa epidemia wirusa Zachodniego Nilu (WNV) dotknęła 71 osób z zapaleniem opon mózgowo-rdzeniowych w Andaluzji (Hiszpania). Próbki od tych osób zostały odebrane przez nasze laboratorium, regionalne Centrum Informacji o Wirusach.
- Celem tego badania było porównanie zgodności, czułości i swoistości wyników między testem WNV VIRCLIA IgG i IgM (Vircell, Hiszpania) a testem WNV ELISA IgM i IgG (Euroimmun, Niemcy) oraz porównanie wydajności WNV VIRCLIA IgM i Euroimmun ELISA do diagnostyki płynu mózgowo-rdzeniowego (CSF).
- Badanie obejmowało 24 próbki płynu mózgowo-rdzeniowego (sparowane z próbkami surowicy) i 247 próbek surowicy od 217 pacjentów z podejrzeniem infekcji WNV (1 lub 2 na pacjenta). Zgodność testów ELISA i CLIA w wykrywaniu IgM i Ig G w surowicy wyniosła odpowiednio 93% (wskaźnik kappa = 0,85) i 96% (wskaźnik kappa = 0,89). Wartości czułości testów ELISA i CLIA dla IgM w próbkach surowicy wyniosły odpowiednio 96,7% i 98,9%, a wartości swoistości odpowiednio 96,4% i 95,4%.
- ll[]l;p[;/.p]lo0Wartości czułości testu ELISA i CLIA dla IgG w próbkach surowicy wyniosły odpowiednio 91,1% i 97%, a wartości swoistości odpowiednio 100% i 98,8%. Wyniki uzyskane za pomocą testów ELISA i CLIA w próbkach płynu mózgowo-rdzeniowego wykazały 75% zgodność między nimi (wskaźnik kappa=0,51). Zgodnie z tymi odkryciami monotest WNV VIRCLIA IgM i IgG zapewnia dokładne jakościowe wykrywanie WNV w próbkach surowicy i płynu mózgowo-rdzeniowego.
W tej pracy pożądane było wprowadzenie nanocząstki polidopaminy (PDA) inspirowanej małżami o biokompatybilnej i reaktywnej charakterystyce powierzchni, charakteryzującej się wysoką nośnością i wywołanym pH zachowaniem uwalniającym jony Fe(III) w celu opracowania bezenzymowego testu kolorymetrycznego.
Katecholowa sieć funkcjonalna PDA może działać jako rusztowanie koordynujące z dużymi ilościami jonów Fe(III) w celu utworzenia NP PDA-Fe(III), których stan koordynacji można dostosować poprzez zmianę wartości pH. W szczególności, NP PDA-Fe(III) można utrzymywać w stanie rozłożonym na trzech współrzędnych w warunkach alkalicznych, podczas gdy silnie obciążone jony Fe(III) mogą być łatwo uwalniane w warunkach kwasowych, aby reagować z żelazocyjankiem w celu wygenerowania in situ pruskiego niebieskie (PB) NP.
Czułość wykrywania antygenu specyficznego dla prostaty (PSA) została znacznie poprawiona dzięki wysokiej aktywności peroksydazy NP PB, która wywołała doskonały efekt katalityczny w reakcji kolorymetrycznej. Ponadto stwierdzono korzystną liniowość w zakresie 0,0005-20 ng mL -1 z dolną granicą wykrywalności 0,84 pg mL -1 (S/N = 3). Ponadto, opracowany bezenzymatyczny test immunologiczny kolorymetryczny wykazał doskonałą selektywność i powtarzalność. Uważa się, że ten proponowany, wolny od enzymów system kolorymetryczny PDA-Fe(III) NP będzie stanowić łatwe i niezawodne narzędzie do czułego wykrywania PSA i innych biomarkerów raka w ludzkiej surowicy.
Jako typowa, klasyczna, ale potężna technologia wykrywania biochemicznego w chemii analitycznej, test immunoenzymatyczny (ELISA) wykazuje doskonałość i szeroką praktyczność w ilościowym oznaczaniu analitów o ultraniskich stężeniach.
However, long incubation time and burdensome laborious multistep washing processes make it inefficient and labor-intensive for conventional ELISA. Here, we propose rod-like magnetically driven nanorobots (MNRs) for use as maneuverable immunoassay probes that facilitate a strategy for an automated and highly efficient ELISA analysis, termed nanorobots enabled ELISA (nR-ELISA). To prepare the MNRs, the self-assembled chains of Fe3O4 magnetic particles are chemically coated with a thin layer of rigid silica oxide (SiO2), onto which capture antibody (Ab1) is grafted to further achieve magnetically maneuverable immunoassay probes (MNR-Ab1s).
We investigate the fluid velocity distribution around the MNRs at microscale using numerical simulation and empirically identify the mixing efficiency of the actively rotating MNRs. To automate the analysis process, we design and fabricate by 3-D printing a detection unit consisting of three function wells. The MNR-Ab1s can be steered into different function wells for required reaction or wishing process.
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16-Channel Fluorescence Immunoassay Analyzer |
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Human Albumin Chemiluminescent Immunoassay Kit |
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Mouse Insulin Chemiluminescent Immunoassay Kit |
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Human Insulin Chemiluminescent Immunoassay Kit |
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Tg ultra-pure IgG low (for Immunoassay use) |
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Rat Mucin 2 Chemiluminescent Immunoassay Kit |
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| IRTMUC2KTC | Innovative research | each | 545 EUR |
Human Aggrecan Chemiluminescent Immunoassay Kit |
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| IHUACANKTC | Innovative research | each | 545 EUR |
Human Arginase Chemiluminescent Immunoassay Kit |
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| IHUARGKTC | Innovative research | each | 545 EUR |
Human Endoglin Chemiluminescent Immunoassay Kit |
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| IHUENGKTC | Innovative research | each | 545 EUR |
Human Ferritin Chemiluminescent Immunoassay Kit |
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| IHUFTKTC | Innovative research | each | 545 EUR |
Human Brevican Chemiluminescent Immunoassay Kit |
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| IHUBCANKTC | Innovative research | each | 545 EUR |
Human Biglycan Chemiluminescent Immunoassay Kit |
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| IHUBGNKTC | Innovative research | each | 545 EUR |
Rat Angiogenin Chemiluminescent Immunoassay Kit |
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| IRTANGKTC | Innovative research | each | 545 EUR |
Rat Calmodulin Chemiluminescent Immunoassay Kit |
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| IRTCAMKTC | Innovative research | each | 545 EUR |
Rat Endostatin Chemiluminescent Immunoassay Kit |
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| IRTCOL18A1KTC | Innovative research | each | 545 EUR |
Rat Calretinin Chemiluminescent Immunoassay Kit |
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| IRTCRKTC | Innovative research | each | 545 EUR |
Rat Calcitonin Chemiluminescent Immunoassay Kit |
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| IRTCTKTC | Innovative research | each | 545 EUR |
Rat Cytovillin Chemiluminescent Immunoassay Kit |
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| IRTCVLKTC | Innovative research | each | 545 EUR |
Rat Dystrophin Chemiluminescent Immunoassay Kit |
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| IRTDMDKTC | Innovative research | each | 545 EUR |
Rat Heparanase Chemiluminescent Immunoassay Kit |
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| IRTHPSEKTC | Innovative research | each | 545 EUR |
Rat Prohibitin Chemiluminescent Immunoassay Kit |
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| IRTPHBKTC | Innovative research | each | 545 EUR |
Rat Proinsulin Chemiluminescent Immunoassay Kit |
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| IRTPIKTC | Innovative research | each | 545 EUR |
Rat Telomerase Chemiluminescent Immunoassay Kit |
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| IRTTEKTC | Innovative research | each | 545 EUR |
Rat Tyrosinase Chemiluminescent Immunoassay Kit |
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| IRTTYRKTC | Innovative research | each | 545 EUR |
Mouse Arginase Chemiluminescent Immunoassay Kit |
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| IMSARGKTC | Innovative research | each | 545 EUR |
Mouse Endoglin Chemiluminescent Immunoassay Kit |
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| IMSENGKTC | Innovative research | each | 545 EUR |
Mouse Ferritin Chemiluminescent Immunoassay Kit |
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| IMSFTKTC | Innovative research | each | 545 EUR |
Mouse Granulin Chemiluminescent Immunoassay Kit |
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| IMSGRNKTC | Innovative research | each | 545 EUR |
Mouse Gelsolin Chemiluminescent Immunoassay Kit |
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| IMSGSKTC | Innovative research | each | 545 EUR |
Mouse Hepcidin Chemiluminescent Immunoassay Kit |
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| IMSHAMPKTC | Innovative research | each | 545 EUR |
Mouse Resistin Chemiluminescent Immunoassay Kit |
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| IMSRETNKTC | Innovative research | each | 545 EUR |
Mouse Visfatin Chemiluminescent Immunoassay Kit |
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| IMSVFKTC | Innovative research | each | 545 EUR |
Mouse Lysozyme Chemiluminescent Immunoassay Kit |
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| IMSLYZKTC | Innovative research | each | 545 EUR |
Human Occludin Chemiluminescent Immunoassay Kit |
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| IHUOCLNKTC | Innovative research | each | 545 EUR |
Human Paxillin Chemiluminescent Immunoassay Kit |
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| IHUPXNKTC | Innovative research | each | 545 EUR |
Human Resistin Chemiluminescent Immunoassay Kit |
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| IHURETNKTC | Innovative research | each | 545 EUR |
Human Survivin Chemiluminescent Immunoassay Kit |
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| IHUSURVKTC | Innovative research | each | 545 EUR |
Human Chemerin Chemiluminescent Immunoassay Kit |
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| IHUTIG2KTC | Innovative research | each | 545 EUR |
Human Vinculin Chemiluminescent Immunoassay Kit |
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| IHUVCLKTC | Innovative research | each | 545 EUR |
Human Visfatin Chemiluminescent Immunoassay Kit |
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| IHUVFKTC | Innovative research | each | 545 EUR |
Human Vimentin Chemiluminescent Immunoassay Kit |
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| IHUVIMKTC | Innovative research | each | 545 EUR |
Human Granulin Chemiluminescent Immunoassay Kit |
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| IHUGRNKTC | Innovative research | each | 545 EUR |
Human Gelsolin Chemiluminescent Immunoassay Kit |
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| IHUGSKTC | Innovative research | each | 545 EUR |
The actively rotating MNR-Ab1s can enhance the binding efficacy with target analytes at microscale and greatly decrease incubation time. The integrated nR-ELISA system can significantly reduce the assay time, more importantly during which process manpower input is greatly minimized. Our simulation of the magnetic field distribution generated by Helmholtz coils demonstrates that our approach can be scaled up, which proves the feasibility of using current strategy to construct high throughput nR-ELISA detection instrument. This work of taking magnetic micro/nanobots as active immunoassay probes for automatic and efficient ELISA not only holds great potential for point-of-care testing (POCT) in future but also extends the practical applications of self-propelled micro/nanorobots into the field of analytical chemistry.