scholarly journals Development of a rapid and sensitive immunochromatographic strip based on EuNPs-ES fluorescent probe for the detection of early Trichinella spiralis-specific IgG antibody in pigs

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Xinyu Wang ◽  
Bin Tang ◽  
Ying Zhao ◽  
Jing Ding ◽  
Nan Wang ◽  
...  

AbstractTrichinellosis, which is caused by nematodes of the genus Trichinella, is one of the most important zoonotic parasite diseases in the world. A rapid and sensitive immunochromatographic strip (ICS) based on Eu (III) nanoparticles (EuNPs) was developed for the detection of Trichinella spiralis (T. spiralis) infection in pigs. T. spiralis muscle larvae excretory secretory or preadult worm excretory secretory (ML-ES or PAW-ES) antigens were conjugated with EuNPs probes to capture T. spiralis-specific antibodies in pig sera, after which the complex bound to mouse anti-pig IgG deposited on the test line (T-line), producing a fluorescent signal. In the pigs infected with 100, 1000 and 10 000 ML, seroconversion was first detectable for the EuNPs-ML-ES ICS at 30, 25 and 21 days post-infection (dpi) and for the EuNPs-PAW-ES ICS at 25, 21 and 17 dpi. These results show that EuNPs-PAW-ES ICS detects anti-Trichinella IgG in pigs 4–5 days earlier that test using ML-ES antigens. Our ICS have no cross reaction with other parasite infection sera. Furthermore, the detection process could be completed in 10 min. This study indicated that our ICS can be used for the detection of the circulating antibodies in early T. spiralis infection and provide a novel method for on-site detection of T. spiralis infection in pigs.

Author(s):  
Cheng-Cheng ZHAI ◽  
Zhao-Jin SUN ◽  
Ming-Yuan LIU ◽  
Xiao-Lei LIU ◽  
Xue BAI ◽  
...  

Background: To assay the Trichinella-specific IgM and IgG antibody responses during the early stage of infection, serum was collected from mice infected with the muscle larvae (ML) of T. spiralis (ISS534) at different dpi (days post infection) up to 60 days. Methods: The levels of IgM and IgG antibodies in serum were measured by ES antigens from different stage of T. spiralis using the ELISA method in Shanghai, China in 2017. Results: The anti-Trichinella IgM and IgG could be detected by ES antigens from the adult three days worm (Ad3) as early as 5 dpi and 8 dpi, respectively. ES antigens from the mixture of adult six days worm & new born larvae (Ad6+NBL) was similar to Ad3. When antibodies were detected by these two antigens, the levels of IgM peaked at 14 dpi and then declined from 15 dpi to 60 dpi; the IgG peaked at 20 dpi, and gradually declined, however, higher detection levels were maintained until 60 dpi. Conclusion: Ad3 ES antigens showed more antigenicity than Ad6+NBL ES on titer detection of IgM and IgG antibodies, and the production of Ad3 ES is easier. In terms of early diagnosis, these two antigens are better than the ML ES antigens of T. spiralis, which antibodies could not be detected before 20dpi. Ad3 ES antigens might be good candidate for the early diagnosis of trichinellosis or the mixture of Ad3 and Ad6+NBL ES might be used.


2019 ◽  
Vol 26 (7) ◽  
pp. 542-549 ◽  
Author(s):  
Shan Shan Hao ◽  
Man Man Zong ◽  
Ze Zhang ◽  
Jia Xi Cai ◽  
Yang Zheng ◽  
...  

Background: Bursa of Fabricius is the acknowledged central humoral immune organ. The bursal-derived peptides play the important roles on the immature B cell development and antibody production. Objective: Here we explored the functions of the new isolated bursal hexapeptide and pentapeptide on the humoral, cellular immune response and antigen presentation to Avian Influenza Virus (AIV) vaccine in mice immunization. Methods: The bursa extract samples were purified following RP HPLC method, and were analyzed with MS/MS to identify the amino acid sequences. Mice were twice subcutaneously injected with AIV inactivated vaccine plus with two new isolated bursal peptides at three dosages, respectively. On two weeks after the second immunization, sera samples were collected from the immunized mice to measure AIV-specific IgG antibody levels and HI antibody titers. Also, on 7th day after the second immunization, lymphocytes were isolated from the immunized mice to detect T cell subtype and lymphocyte viabilities, and the expressions of co-stimulatory molecule on dendritic cells in the immunized mice. Results: Two new bursal hexapeptide and pentapeptide with amino acid sequences KGNRVY and MPPTH were isolated, respectively. Our investigation proved the strong regulatory roles of bursal hexapeptide on AIV-specific IgG levels and HI antibody titers, and lymphocyte viabilities, and the significant increased T cells subpopulation and expressions of MHCII molecule on dendritic cells in the immunized mice. Moreover, our findings verified the significantly enhanced AIV-specific IgG antibody and HI titers, and the strong increased T cell subpopulation and expressions of CD40 molecule on dendritic cells in the mice immunized with AIV vaccine and bursal pentapeptide. Conclusion: We isolated and identified two new hexapeptide and pentapeptide from bursa, and proved that these two bursal peptides effectively induced the AIV-specific antibody, T cell and antigen presentation immune responses, which provided an experimental basis for the further clinical application of the bursal derived active peptide on the vaccine improvement.


2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Ge Li ◽  
Xun Wang ◽  
Qingmei Li ◽  
Jifei Yang ◽  
Xiao Liu ◽  
...  

Abstract Background H7N9 avian influenza virus (AIV) including highly and low pathogenic viruses have been detected in China since 2013. H7N9 AIV has a high mortality rate after infection in humans, and most human cases have close contacted with poultry in the live poultry market. Therefore, it is necessary to develop a rapid point-of-care testing (POCT) technique for H7N9 AIV detection. Methods The H7N9 AIV was inactivated and purified, and was used as the antigen to immunize BALB/c. Twelve H7-HA specific monoclonal antibodies (McAbs) were produced through the hybridoma technique. The McAb 10A8 was conjugated with colloid gold as detecting antibody; McAb 9B6 was dispensed on the nitrocellulose membran as the capture test line and the Goat-anti mouse IgG antibody was dispensed as control line respectively. The immunochromatographic strip was prepared. Results The analysis of ELISA and virus neutralization test showed that the obtained McAbs specifically recognized H7 HA. Based on the prepared strip, the detection of H7 AIV was achieved within 10 min. No cross-reaction occurred between H7 AIVs and other tested viruses. The detection limit of the strip for H7 was 2.4 log10EID50/0.1 mL for chicken swab samples. Conclusion The McAbs were specific for H7 and the immunochromatographic strip developed in this study was convenient, rapid and reliable for the detection of H7 AIV. The strip could provide an effective method for the rapid and early detection of H7 AIV.


2002 ◽  
Vol 88 (11) ◽  
pp. 1004-1007 ◽  
Author(s):  
Alisa Gruden-Movsesijan ◽  
Natasa Ilic ◽  
Ljiljana Sofronic-Milosavljevic

2005 ◽  
Vol 66 (8) ◽  
pp. 2
Author(s):  
Dessislava Kopchaliiska ◽  
Mary S. Leffell ◽  
Andrea A. Zachary

1980 ◽  
Vol 85 (3) ◽  
pp. 381-391 ◽  
Author(s):  
J. E. Cradock-Watson ◽  
Margaret K. S. Ridehalgh ◽  
Mary J. Anderson ◽  
J. R. Pattison ◽  
H. O. Kangro

SUMMARYWe have tried to measure the incidence of prenatal infection in 304 infants whose mothers had had rubella at various times after the first 12 weeks of pregnancy. Two methods of assessment were used: first, serum obtained soon after birth was tested for specific IgM antibody; secondly, serum obtained after the age of eight months was tested for specific IgG. When maternal rubella occurred 12–16 weeks after the last menstrual period specific IgM antibody was detected in 28 out of 50 infants (56%). The proportion fell progressively to 12% after maternal rubella at 24–28 weeks, rose to 19% after rubella at 28–36 weeks and then to 58% when the illness occurred during the last month of pregnancy. In all, IgM antibody was detected in 77 out of 260 infants (29%). The fetus can thus be infected at any time during the second and third trimesters of pregnancy, but the risk varies at different stages.The figures for the prevalence of IgG antibody were greater throughout, because some infants had IgG who had previously lacked specific IgM. After maternal rubella at 12–16 weeks IgG antibody persisted in 22 out of 31 infants (71%). The proportion fell to 28% after rubella at 24–28 weeks and then increased progressively to 94% after rubella during the last month. In all, IgG antibody persisted in 94 out of 190 infants (49%). The true rate of fetal infection probably lies between the rates estimated from the presence of IgM antibody and the subsequent prevalence of IgG.Infants whose mothers had rubella at any time during pregnancy should be examined regularly for possible evidence of damage.


1986 ◽  
Vol 79 (4) ◽  
pp. 344-348 ◽  
Author(s):  
D. Lee ◽  
Janet M. Dewdney ◽  
R.G. Edwards ◽  
K.A. Neftel ◽  
M. Wälti

Parasitology ◽  
2010 ◽  
Vol 138 (4) ◽  
pp. 463-471 ◽  
Author(s):  
ALEKSANDAR ZOCEVIC ◽  
PAULINE MACE ◽  
ISABELLE VALLEE ◽  
RADU BLAGA ◽  
MINGYUAN LIU ◽  
...  

SUMMARYThree expression cDNA libraries fromTrichinella spiralisworms 14 h, 20 h and 48 h post-infection (p.i.) were screened with serum from pigs experimentally infected with 20 000T. spiralismuscle larvae. Twenty-nine positive clones were isolated from the 14 h p.i. cDNA library, corresponding to 8 different genes. A putative excretory-secretory protein similar to that ofT. pseudospiraliswas identified. Three clones corresponded to aT. spiralisserine proteinase inhibitor known to be involved in diverse functions such as blood coagulation and modulation of inflammation. Screening of the 20 h p.i. cDNA library selected 167 positive clones representing 12 different sequences. The clone with the highest redundancy encoded a small polypeptide having no sequence identity with any known proteins fromTrichinellaor other organisms. Fourteen clones displayed sequence identity with the heat shock protein (HSP) 70. HSPs are produced as an adaptive response of the parasite to the hostile environment encountered in the host intestine but their mechanism of action is not yet well defined. From the 48 h p.i.T. spiraliscDNA library, 91 positive clones were identified representing 7 distinct sequences. Most of the positive clones showed high similarity with a member of a putativeT. spiralisserine protease family. This result is consistent with a possible major role for serine proteases during invasive stages ofTrichinellainfection and host-parasite interactions.


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