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Background: Non-structural glycoprotein-1 (NS1) is a useful biomarker for early diagnosis of dengue fever. NS-1 antigen ELISA can be used for the early diagnosis of dengue fever in the acute stage. Quantitative methods are better for epidemic settings due to high false negative rates in qualitative ELISA. Methods: The study was initiated after approval from the institutional ethics council (IEC/DISS/17118). Study examined 280 patients with dengue symptoms who presented to the hospital's OPDs and IPDs. Patients were tested using qualitative ELISA, and those with Leptospira antibody, malaria, or Chikungunya IgM antibody were excluded. Age, gender, symptoms, comorbidities, total leucocyte count, platelet count, and risk category were all patient-related parameters. Patient-related parameters were recorded, and data was collected using Microsoft excel and analysed statistically. Results: Most patients aged 2-40 with male predominance had fever, chills, and body aches, 243 (86.8%) tested positive for ELISA NS1. Quantitative ELISA test showed a statistically significant correlation with rapid antigen NS1 result (p=0.015). Its AUC was 0.883 (p=0.0001), and its cut-off was (>109.1) with 96.9% sensitivity and 13.64% specificity. The AUC of quantitative ELISA NS1 against qualitative ELISA NS1 was 0.853 which was statistically significant (p<0.0001). At the cut-off >74.34, the test's sensitivity was 92.59% and specificity was 75.68%. Conclusions: Qualitative ELISA NS1 test is better than rapid antigen test for screening due to its higher specificity and similar sensitivity.
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@#Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 × 10~5 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines
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@#Objective To prepare rabbit polyclonal antibodies against pertussis toxin(PT) and develop a double antibody sandwich ELISA for quantitative determination of PT antigen,identify and apply the method.Methods The rabbit polyclonal antibody against PT was prepared by immunizing Chinchilla rabbit with PT using traditional method.The reaction conditions of ELISA system were optimized,the double antibody sandwich ELISA method for quantitative determi-nation of PT was developed,and the specificity,linearity,accuracy,precision and sensitivity were verified.The developed method was used to detect PT antigen content in fimbriae proteins(FIM) stock solution of samples during detoxification and other purification process of pertussis antigen.Results The working condition of double antibody sandwich ELISA for detection of PT antigen content was the coating concentration of PT rabbit polyclonal antibody of 1 μg/mL,and the enzyme-labeled antibody dilution of 1:8 000.This detection system showed specific reaction with PT purified protein,but had no cross reaction with filamentous hemagglutinin,diphtheria toxoid and tetanus toxoid;the linear detection range of the developed double antibody sandwich ELISA was within 25—400 ng/mL;the recovery rates of PT at high,moderate and low concentrations were 103.27%,91.48% and 103.52%,respectively;both the intra-and inter-coefficients of variation(CVs)were less than 10%;the sensitivity of the method was 20.719 ng/mL,and the detection limit was 41.438 ng/mL.Thirty-five batches of samples were detected under five different detoxification process conditions and at different sampling time points,and the changes of antigen content were all consistent with the trend of detoxification reaction.Conclusion The PT rabbit polyclonal antibody was successfully prepared,and a double antibody sandwich ELISA with high precision and accuracy was developed for the quantitative determination of PT antigen content,which can be used for the antigen content detection of chemically detoxified samples in the production process of component DPT vaccines.
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Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.
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@#Objective To establish and validate a method for the determination of the interesting protein expression level of recombinant adeno-associated virus(rAAV)infected cells,so as to monitor the product quality in different stages of rAAV9production process.Methods After incubation of serial diluted rAAV samples with infection enhancer Envirus-AAV,the human malignant glioblastoma cells(U87-MG)pretreated with hydroxyurea(HU)were infected.Using rAAV9 reference as the standard,the expression level of glutaryl-CoA dehydrogenase(GCDH)was detected by ELISA,and the specificity,accuracy,precision,linear range,limit of quantitation(LOQ)and durability of the method were verified.Eight batches of rAAV9 samples were detected by the established method.Results The A_(450)-A_(630) value of the sample buffer was 0.3,which was slightly lower than the lowest dilution point(1 ng/mL)of the four-parameter standard curve for protein quantification.The average recoveries of samples with 150%,100% and 50% theoretical relative titer levels were in the range of 100.0%-107.3%.The RSDs of the target protein expression level of the samples with three theoretical relative titer levels detected by the same experimenter three times and different experimenters were all less than 25%.There was a good linear relationship between rAAV9 samples and the target protein expression levels in the range of 50%-150% theoretical relative titer levels,and the linear regression equation was y = 1.077 x-0.022,R~2= 0.984.The LOQ of the method was 0.59,namely 6.0×10~(12) vg/mL.After U87-MG cells were incubated with HU for different time(18,21,24 h),and the culture supernatant was stored under different conditions(room temperature for 0.5 h,below-60 ℃ for 12 h,below-60 ℃ for 24 h).The RSDs of target protein expression levels were all less than 25%.The target protein expression levels of 1-8 batches of rAAV9 samples were 111%,121%,72%,65%,86%,75%,102% and 91%,respectively.Conclusion The established method for the determination of the target protein expression level after rAAV infection has good specificity,accuracy,precision and durability,and can be used for the quality control of products in different stages of rAAV9 production.
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【Objective】 To explore the epidemiological characteristics of voluntary blood donors with enzyme-linked immuno-sorbent assay (ELISA) negative and nucleic acid testing (NAT) positive in Hainan from 2012 to 2022, so as to provide reference for developing rational blood screening strategies. 【Methods】 The screening results for transfusion-transmitted disease markers in 1 161 042 blood samples in Hainan from 2012 to 2022 were retrospectively analyzed. All samples have been measured twice by ELISA and once by NAT. Statistical methods were used to analyze the proportion of ELISA negative and NAT positive (ELISA-/NAT+ ) among voluntary blood donors and its relation with factors including gender, age, ethnicity and region. 【Results】 Among the voluntary blood donors in Hainan from 2012 to 2022, the overall proportion of ELISA-/NAT+ was 0.19% (2 151/1 161 042), and the difference was statistically significant (P<0.05). The ELISA-/NAT+ rate in hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA, human immunodeficiency virus (HIV) RNA and non-discriminating reactive (NDR) was 0.10%, 0.000 3%, 0.000 4% and 0.09% respectively. The ELISA-/NAT+ rate of voluntary blood donors varied among different age groups and gradually increased with age (P<0.05). The ELISA-/NAT+ rate of male donors (0.22%, 1 729/795 032) was significantly higher than that of female donors (0.12%, 422/366 010, P<0.05). The ELISA-/NAT+ rate of Han blood donors was significantly lower than that of Li and Miao blood donors (P<0.05). The ELISA-/NAT+ rate was the highest of 0.32% (301/94 046) in the eastern region, followed by 0.30% (341/113 783) in western region, and 0.16% in both southern and northern region, which also presented a significant difference (P<0.05). 【Conclusion】 The ELISA-/NAT+ rate of voluntary blood donors in Hainan fluctuated from 2012 to 2022, which was related to factors such as age, gender, ethnicity and region.
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【Objective】 To analyze the effect of sample hemolysis on ELISA test results in blood screening laboratory, so as to determine the acceptable tolerance of hemolysis specific to laboratory test items and detection system, and provide reference for the formulation of tolerance standard of sample hemolysis. 【Methods】 Negative and weakly positive (S/CO was about 2) samples with different hemolysis degrees were tested by several commonly used domestic reagents for HBsAg, HIV Ag/Ab, anti-HCV and anti-TP, respectively. The effects of various degrees of hemolysis on the test results of negative and weakly positive samples for each item were analyzed. 【Results】 1) Hemolysis had no effect on the test results (reactive/non-reactive) of negative and weakly positive samples for HBsAg, anti-HCV and anti-TP ELISA items; 2) Hemolysis affected the test results (reactive/non-reactive) of negative and weakly positive samples for HIV Ag/Ab ELISA item. A tolerance of Hb 2 g/L was taken as the acceptable hemolysis degree for HIV Ag/Ab ELISA item. 【Conclusion】 In this study, the acceptable tolerance of hemolytic samples for corresponding test items and detection system in our laboratory were determined. The influence of hemolysis on ELISA test result is related to the reagent, equipment, environment and other factors, therefore the acceptable tolerance of hemolysis should be determined scientifically and reasonably based on the specific evaluation of each laboratory.
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@#Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 × 10~5 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines.
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@#Objective To prepare rabbit polyclonal antibodies against pertussis toxin(PT) and develop a double antibody sandwich ELISA for quantitative determination of PT antigen,identify and apply the method.Methods The rabbit polyclonal antibody against PT was prepared by immunizing Chinchilla rabbit with PT using traditional method.The reaction conditions of ELISA system were optimized,the double antibody sandwich ELISA method for quantitative determi-nation of PT was developed,and the specificity,linearity,accuracy,precision and sensitivity were verified.The developed method was used to detect PT antigen content in fimbriae proteins(FIM) stock solution of samples during detoxification and other purification process of pertussis antigen.Results The working condition of double antibody sandwich ELISA for detection of PT antigen content was the coating concentration of PT rabbit polyclonal antibody of 1 μg/mL,and the enzyme-labeled antibody dilution of 1:8 000.This detection system showed specific reaction with PT purified protein,but had no cross reaction with filamentous hemagglutinin,diphtheria toxoid and tetanus toxoid;the linear detection range of the developed double antibody sandwich ELISA was within 25—400 ng/mL;the recovery rates of PT at high,moderate and low concentrations were 103.27%,91.48% and 103.52%,respectively;both the intra-and inter-coefficients of variation(CVs)were less than 10%;the sensitivity of the method was 20.719 ng/mL,and the detection limit was 41.438 ng/mL.Thirty-five batches of samples were detected under five different detoxification process conditions and at different sampling time points,and the changes of antigen content were all consistent with the trend of detoxification reaction.Conclusion The PT rabbit polyclonal antibody was successfully prepared,and a double antibody sandwich ELISA with high precision and accuracy was developed for the quantitative determination of PT antigen content,which can be used for the antigen content detection of chemically detoxified samples in the production process of component DPT vaccines
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@#Objective To develop and verify a double-antibody sandwich ELISA method for the detection of process-specific E.coli residual protein in recombinant biological preparations.Methods Taking the production and purification process of glucagon-like peptide(GLP)expressed by E.coli as the specific process model,the same process was used to intercept the residual protein of empty E.coli(normal E.coli that does not express recombinant protein). One female New Zealand white rabbit and six female Kunming mice were immunized with the residual protein as the immunogen. Using the IgG antibody purified from rabbit immune serum as the coating antibody,mouse immune serum as the second sandwich antibody,and antimouse IgG-HRP as the enzyme-labeled secondary antibody,a double antibody sandwich ELISA method for process-specific residual protein of E.coli was established. The specificity,accuracy and precision of the method were verified,and the limit of detection(LOD)was determined. Simultaneously,the developed method and the commercial E.coli host protein residue detection kit were used to quantitatively determine the residual protein of purified GLP preparation.Results After a series of gradient dilution of process-specific residual protein with known concentration,the sensitivity of this ELISA method reached 338 pg/mL. No cross reaction occurred in the detection of CHO and yeast cell lysis protein by this method,the recoveries of samples with low,medium and high concentrations were all in the range of 80% — 120%,and the intra-assay and inter-assay CVs of the empty E.coli interception standard with low,medium and high concentrations were all less than15%. For the residual protein in GLP preparation,about 62% of the residual proteins were not detected by the commercial non-process-specific ELISA kit compared with the total amount of residual proteins detected by the developed method,and these residual proteins should be the process-specific residual proteins.Conclusion The double antibody sandwich ELISA method developed in this study has high sensitivity,strong specificity,good accuracy and precision for the detection of process-specific E.coli residual protein,which can meet the detection requirements that the residual protein is less than0. 01% — 0. 1% in biological preparations.
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Local and exotic germplasm of tomato remains a major source for genetic improvement. Assessment of such lines for biotic stresses particularly viral diseases are the most important criteria for selection in Pakistan, where Tomato Yellow Leaf Curl Virus (TYLCV) and Tomato Mosaic Virus (ToMV) are the major diseases/viruses. A set of 40 accessions (including indigenous Pakistani lines and exotic germplasm from Europe, the United States, and Asia) were evaluated for their resistance/infection response to ToMV with artificial inoculation under greenhouse conditions. Infection response was quantified through disease scoring and DAS-ELISA test (for ToMV). A subset of 24 lines, was further screened for TYLCV using disease scoring and TAS-ELISA. The tested lines showed significant variability for resistance to ToMV. Only one accession (Acc-17878) was resistant to the ToMV whereas seven accessions i.e. Acc-17890, AVR-261, CLN-312, AVR-321, EUR-333, CLN-352, and CLN-362 expressed resistance to TYLCV. Correlation between phenotypic evaluation was confirmed by the ELISA results in both diseases, although both tools complemented to assess the viral infection status. In future, tomato breeding programs must consider breeding for ToMV and TYLCV resistance (using identified germplasm in our study) so as to deliver virus resistant tomato varieties.
O germoplasma local e exótico do tomate continua sendo uma importante fonte de melhoramento genético. A avaliação de linhagens para estresses bióticos, particularmente as doenças virais, é o critério mais importantes para seleção no Paquistão, onde o vírus da folha amarela do tomate (TYLCV) e o vírus do mosaico do tomateiro (ToMV) são as principais doenças/vírus. Um conjunto de 40 acessos (incluindo linhagens indígenas do Paquistão e germoplasma exótico da Europa, dos Estados Unidos e da Ásia) foi avaliado quanto à resistência/resposta à infecção ao ToMV com inoculação artificial em casa de vegetação. A resposta à infecção foi quantificada por meio de pontuação da doença e de teste DAS-ELISA (para ToMV). Um subconjunto de 24 linhas foi posteriormente rastreado para TYLCV usando pontuação de doença e TAS-ELISA. As linhas testadas apresentaram variabilidade significativa para resistência ao ToMV. Apenas um acesso (Acc-17878) foi resistente ao ToMV, enquanto sete acessos (Acc-17890, AVR-261, CLN-312, AVR-321, EUR-333, CLN-352 e CLN-362) expressaram resistência ao TYLCV. A correlação entre a avaliação fenotípica foi confirmada pelos resultados do ELISA nas duas doenças, embora ambas as ferramentas tenham se complementado para avaliar o estado da infecção viral. No futuro, os programas de melhoramento de tomate devem considerar aperfeiçoamentos para resistência ao ToMV e TYLCV (usando germoplasma identificado em nosso estudo) de modo a fornecer variedades de tomate resistentes a vírus.
Subject(s)
Solanum lycopersicum , Genetic Enhancement , Mosaic VirusesABSTRACT
Abstract Local and exotic germplasm of tomato remains a major source for genetic improvement. Assessment of such lines for biotic stresses particularly viral diseases are the most important criteria for selection in Pakistan, where Tomato Yellow Leaf Curl Virus (TYLCV) and Tomato Mosaic Virus (ToMV) are the major diseases/viruses. A set of 40 accessions (including indigenous Pakistani lines and exotic germplasm from Europe, the United States, and Asia) were evaluated for their resistance/infection response to ToMV with artificial inoculation under greenhouse conditions. Infection response was quantified through disease scoring and DAS-ELISA test (for ToMV). A subset of 24 lines, was further screened for TYLCV using disease scoring and TAS-ELISA. The tested lines showed significant variability for resistance to ToMV. Only one accession (Acc-17878) was resistant to the ToMV whereas seven accessions i.e. Acc-17890, AVR-261, CLN-312, AVR-321, EUR-333, CLN-352, and CLN-362 expressed resistance to TYLCV. Correlation between phenotypic evaluation was confirmed by the ELISA results in both diseases, although both tools complemented to assess the viral infection status. In future, tomato breeding programs must consider breeding for ToMV and TYLCV resistance (using identified germplasm in our study) so as to deliver virus resistant tomato varieties.
RESUMO O germoplasma local e exótico do tomate continua sendo uma importante fonte de melhoramento genético. A avaliação de linhagens para estresses bióticos, particularmente as doenças virais, é o critério mais importantes para seleção no Paquistão, onde o vírus da folha amarela do tomate (TYLCV) e o vírus do mosaico do tomateiro (ToMV) são as principais doenças/vírus. Um conjunto de 40 acessos (incluindo linhagens indígenas do Paquistão e germoplasma exótico da Europa, dos Estados Unidos e da Ásia) foi avaliado quanto à resistência/resposta à infecção ao ToMV com inoculação artificial em casa de vegetação. A resposta à infecção foi quantificada por meio de pontuação da doença e de teste DAS-ELISA (para ToMV). Um subconjunto de 24 linhas foi posteriormente rastreado para TYLCV usando pontuação de doença e TAS-ELISA. As linhas testadas apresentaram variabilidade significativa para resistência ao ToMV. Apenas um acesso (Acc-17878) foi resistente ao ToMV, enquanto sete acessos (Acc-17890, AVR-261, CLN-312, AVR-321, EUR-333, CLN-352 e CLN-362) expressaram resistência ao TYLCV. A correlação entre a avaliação fenotípica foi confirmada pelos resultados do ELISA nas duas doenças, embora ambas as ferramentas tenham se complementado para avaliar o estado da infecção viral. No futuro, os programas de melhoramento de tomate devem considerar aperfeiçoamentos para resistência ao ToMV e TYLCV (usando germoplasma identificado em nosso estudo) de modo a fornecer variedades de tomate resistentes a vírus.
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Equine influenza is a highly contagious viral disease, specially among 1-5 years old naive horses. Vaccination is considered the best way to control the disease spread and outbreaks. Although foals are the main animal used for evaluation of equine influenza vaccines, guinea pigs were chosen as an alternative model in the present work, as they have a negligible antibody titer against equine influenza virus and are cheaper and easier to handle than foals. Five equine influenza vaccine batches were evaluated in two animal models, foals and guinea pigs, by injection of two doses/animal with 4 weeks apart using 2 mL/animal/dose and evaluation of immune responses by hemagglutination inhibition test and enzyme-linked immunosorbent assay. On the 7th week post vaccination, equine influenza antibodies titers reached maximum values of 9-10.2 and 8.7-10 hemagglutination inhibition units for foals and guinea pigs, respectively; sample/negative ratios were 0.126-0.464 and 0.128-0.445 for both animals, respectively. The use of guinea pigs as an animal model for the evaluation of equine influenza vaccines could be recommended instead of foals(AU)
La gripe equina es una enfermedad viral muy contagiosa, especialmente entre los caballos jóvenes de 1 a 5 años de edad. La vacunación se considera la mejor forma de controlar la propagación y los brotes de la enfermedad. Aunque los potros son el principal animal utilizado para la evaluación de vacunas contra la gripe equina, en el presente trabajo se eligieron cobayos como modelo alternativo, ya que tienen un título insignificante de anticuerpos contra el virus de la gripe equina y son más baratos y fáciles de manejar que los potros. Se evaluaron cinco lotes de vacunas contra la gripe equina en dos modelos animales, potros y cobayos, mediante la inyección de dos dosis/animal con 4 semanas de intervalo utilizando 2 mL/animal/dosis y la evaluación de las respuestas inmunitarias mediante la prueba de inhibición de la hemaglutinación y el ensayo inmunoenzimático. En la 7ª semana posvacunación, los títulos de anticuerpos contra la gripe equina alcanzaron valores máximos de 9-10,2 y 8,7-10 unidades de inhibición de la hemaglutinación para potros y cobayos, respectivamente; las relaciones muestras/negativos fueron de 0,126-0,464 y 0,128-0,445 para ambos animales, respectivamente. Podría recomendarse el uso de cobayos como modelo animal para la evaluación de vacunas contra la gripe equina, en lugar de potros(AU)
Subject(s)
AnimalsABSTRACT
Background: Common antigenic pool is seen because of shared embryonic origins of gall bladder cancer (GBC) and pancreas. Hence, we analyzed the role of serum carbohydrate antigen 242 (CA242) which has been studied in pancreatic cancer, in GBC. The objectives were to identify whether serum CA242 provides added advantage in diagnosis of GBC when compared to controls and to determine its cut-off value. Methods: Serum CA 19-9 level was determined by chemiluminescent micro particle assay and CA242 by enzyme linked immunosorbent assay (ELISA) of age matched cases and controls. Results: Total enrolled patients were 83 including 10 (11.7%) healthy volunteers, 22 (25.9%) chronic cholecystitis cases, and 53 (62.4%) patients with histological evidence of carcinoma. Mean age of presentation of GBC was 51.64 SD10.88 years with F: M ratio of 5.6:1. Pain (90.6%, 48/53) accompanied with jaundice was significantly associated with GBC well reflected by significantly raised serum total bilirubin (p=0.011), direct bilirubin (p=0.008) along with alkaline phosphatase levels (p=0.001). Significantly higher median value of CA 19-9 and CA242 was observed in GBC when compared to CC and healthy volunteers (p<0.001) with a significant correlation between tumor size (>2.5 cm) and serum levels of CA242. The best cut-off limit for CA242 was 45.25 IU/ml. The specificity for carcinoma diagnosis increased to 100% when CA242 was included along with CA 19.9 in serological estimation. Conclusions: We recommend that CA antigen 19-9 may be complimented with CA242 for serological identification of malignancy in the gall bladder.
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Objectives:As a part of an ongoing research programme on vector-borne viral diseases especially Dengue, a retrospective analytical study on the occurrence and distribution of Dengue virus serotype(s) in the state of Manipur – a small state situated in the northeastern region of the Indian subcontinent was carried out at Viral Research & Diagnostic Laboratory (VRDL), Department of Microbiology, Regional Institute of Medical Sciences (RIMS), Imphal which is a tertiary care hospital.Materials & Methods:A total of 914 blood samples from clinically dengue-suspected patients were screened for the presence of Dengue infection by adopting the ELISA (IgM) technique during the period from 01/06/2022 to 02/12/2022. Further, anti-Dengue IgM antibody-positive samples having high Optical Density (OD) value(s) were selected and subjected to RT PCR to determine the serotype(s) of the Dengue virus.Results:Of the 914 blood samples examined for the presence of Dengue infection, 111 (12.14%) were found positive for anti- Dengue IgM antibody indicating acute infection of Dengue virus.Of the positive patients, there were 56 (50.45%) males and 55 (49.54%) females.Predominant clinical features observed among the Dengue-confirmed patients included – fever (74%), headache (19%), arthralgia / joint pain (9%), myalgia (6%) and vomiting (6%) respectively.The study revealed that while the circulation of three (03) Dengue virus serotypes, namely DENV – 1, DENV – 2 & DENV – 3 were observed in the Tengnoupal district,the circulation of two Dengue virus serotypes i.e., DENV – 1 & DENV – 2 was evident in Imphal West & Bishnupur districts respectively.The present study also reveals the occurrence of Dengue virus serotype – 1 (DENV - 1) in Churachandpur district. Conclusion:The present study reveals the circulation/distribution of three Dengue virus serotypes namely, DENV – 1, DENV – 2 and DENV – 3 among the studied samples.
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Introducción . La toxoplasmosis es una zoonosis parasitaria que compromete al ser humano y muchas otras especies de vertebrados, provocada por el agente etiológico Toxoplasma gondii. Objetivo . El objetivo del presente estudio es determinar la frecuencia de toxoplasmosis en muestras de pacientes atendidos en el Instituto SELADIS (*) durante el período de enero de 2021 a julio de 2022, además de su relación con el diagnóstico clínico que incluyen las solicitudes de pruebas. Materiales y métodos . Se consideraron 290 pruebas de pacientes con requerimiento de anticuerpos IgG e IgM anti-Toxoplasma gondii, utilizando la prueba de ELISA (comercial). Resultados . Se encontró que la frecuencia de anticuerpos IgM contra T. gondii fue del 7.24 % (21/290) y de anticuerpos IgG contra T. gondii, del 33.1 % (96/290). En relación con la edad, se observó que en pacientes adultos (mayores de 31 años) la frecuencia fue mayor (IgM: 4.14% % e IgG: 23.8%). El 6.9% de los pacientes tenían un diagnóstico relacionado con patología ocular, el 6.21% eran mujeres embarazadas y entre ellas la seropositividad para anticuerpos IgG fue del 24.64%; y para IgM fue del 8.70% (primer trimestre 2.90% y segundo 4.35%). La seropositividad para anticuerpos IgG, en pacientes trasplantados renales, fue del 1.4% y para IgM del 0%. En pacientes con adenomegalia, el 2.4% fueron positivos para IgG y el 0.69% para IgM. Conclusión . En conclusión, se encontró que la frecuencia de anticuerpos contra toxoplasmosis en la población de estudio fue de 33.1% para IgG y 7.24% para IgM. En relación con el diagnóstico clínico, se encontró que los tres principales escenarios de salud son (en orden descendente de importancia) para el Ac IgG contra toxoplasmosis: patología ocular, el embarazo y adenomegalias. En cambio, la relación con el diagnóstico de estos tres escenarios para el Ac IgM contra toxoplasmosis, son: el embarazo, patología ocular y adenomegalias.
Introduction . Toxoplasmosis is a parasitic zoonosis disease that affects humans and many other vertebrate species, and is caused by the etiological agent known as Toxoplasma gondii. Objective . The aim of this study is to determine the frequency of toxoplasmosis in samples from patients who were assisted in the SELADIS Institute (*) in the period between January 2021 and July 2022, in addition to its relation to the clinical diagnosis including requested tests. Materials and methods . 290 samples from patients were considered that also required IgG and IgM anti-Toxoplasma gondii antibodies, using an ELISA assay (commercial kit). Results. The frequency of IgM antibodies against T. gondii was 7.24% (21/290) and IgG antibodies against T. gondii were 33.1% (96/290). In relation to age, it was observed that in adult patients (over 31 years of age) the frequency was higher (IgM: 4.14%, IgG: 23.8%). 6.9% of the patients had a diagnosis related to ocular pathology, 6.21% were pregnant women and among them the seropositivity for IgG antibodies was 24.64%; and for IgM it was 8.70% (first trimester 2.90% and second 4.35%). The seropositivity for IgG antibodies in kidney transplant patients was 1.4% and 0% for IgM. In patients with adenomegaly, 2.4% were positive for IgG and 0.69% for IgM. Conclusion . In conclusion, it was found that the frequency of antibodies against Toxoplasmosis in the study population was 33.1% for IgG and 7.24% for IgM. In relation to clinical diagnosis, it was found that the three main health scenarios are (in descending order of importance), for IgG Ab against toxoplasmosis: ocular pathology, pregnancy and adenomegaly. Besides this, it was found that the relation of these three scenarios diagnoses to IgM Ab against toxoplasmosis are: pregnancy, ocular pathology and adenomegaly.
Subject(s)
ToxoplasmosisABSTRACT
Scrub typhus is a mite-borne typhus caused by Orientia tsutsugamushi and it is characterized by acute febrile illness, rash, eschar and an incubation period ranging from 6 to 21 d. It affects various organs such as lungs, heart, spleen, liver, and kidney. We report a case on 5 y old male child was admitted with complaints of fever and vomiting. Based on a general examination, the patient had eschar on the scrotum. Diagnosis was made based on clinical features, and the serology IgM for scrub typhus was positive. He was treated with doxycycline. To prevent complications, the patient needs effective management, early administration of antibiotics and preventive measures to control vector bite.
ABSTRACT
Background: Dexamethasone is a synthetic corticosteroid similar to cortisol produced naturally by the adrenal glands. As an anti- inflammatory and immunosuppressive agent, it is used in many diseases such as rheumatoid arthritis and allergic anaphylactic shock, and its suppression test to diagnose Cushing's syndrome. Its further use includes its administration before antibiotics in bacterial meningitis, antitumor treatment, for treatment of glucocorticoid resistance, Addison抯 disease, and congenital adrenal hyperplasia. The drug is abused by using it in animal husbandry as a growth promoter and in horse sports to enhance their performance. Methods: In this study, the development of homologous ELISA using Dexamethasone-21-hemisuccinate (DEX-21-HS)-Bovine serum albumin antiserum and Dexamethasone-21-hemisuccinate (DEX-21-HS)-Horseradish peroxidase enzyme conjugate has been done. The n-hydroxysuccinimide ester method was used to prepare the immunogen and enzyme conjugate. Results: The sensitivity 0.25 ng/mL, affinity 2.8x10-8 L/mol and ED50 4.98 ng/mL of the assay were found. The cross-reactivity of the assay was checked and found with three steroids (Corticosterone- 1.13%, Progesterone- 2.25% and Prednisolone- 6.3%) out of 48 structurally related steroids. Then, analytical variables of the developed assay were studied, such as recovery (98.55% to 105.08%), precision (Inter and Intra- assay coefficient of variation <9.28%), correlation (R2= 0.98) by utilizing a commercially available Dexamethasone kit for comparison. Conclusion: This study concluded that low-cost indigenous ELISA for Dexamethasone had been developed, which can give results within 75-80 minutes.
ABSTRACT
Scrub typhus, a zoonosis, caused by Orientia tsuttsutgamushi is transmitted to man through bite of infected tromboculid mite. It is endemic in India as other other South-east Asia countries of Tsuttsutgamuhi triangle and affects children as well. It generally presents as an acute febrile illness with non-specific features like headache rash, lymphadenopathy resembling many other commonly prevalent febrile infections. An eschar, though pathognomonic of Scrub typhus, is often missed clinically. Therefore, a high index of clinical suspicion is essential to establish a diagnosis of Scrub typhus and hence it remains usually under-diagnosed in any undifferentiated febrile illness. In this case series including three cases, we decided to rule out Scrub typhus as a co-infection when there was persistence of fever associated with thrombocytopenia even after ongoing treatment of the primarily diagnosed infection. An eschar was incidentally detected in one case only.
ABSTRACT
Background: Membranous nephropathy (MN) is a pattern of glomerular injury. Exact categorization into primary membranous nephropathy (PMN) or secondary membranous nephropathy (SMN) is essential for treatment. An endogenous podocyte antigen, M-type phospholipase A2 receptor (PLA2R) has been discovered to be involved in the pathogenesis of PMN. Aims and Objectives: In this article, we aimed to analyze renal tissue PLA2R and serum anti-PLA2R antibodies in MN cases and determined the diagnostic utility. Materials and Methods: The study was of prospective type carried out from March 2019 to August 2020. Analysis of cases of MN was performed with PLA2R paraffin immunoflourescence and serum anti-PLA2R antibody ELISA. Results: Overall sensitivity, specificity, PPV, and NPV of serum anti-PLA2R ELISA for PMN was 91.3%, 80%, 75%, and 93.3%, respectively, and of tissue PLA2R staining for PMN was 91.67%, 81.08%, 75.86%, and 93.75%, respectively. There was strong concordance between two methods. In the patients that were followed up, we found baseline serum anti-PLA2R antibody was less in complete remission group than that in non-remission group and the reduction in serum anti-PLA2R antibody was more in complete remission group than that in non-remission group. Conclusion: Routine light and immunofluorescence examination are incapable of giving exact categorical opinion regarding PMN and SMN. Serum anti-PLA2R antibody detection and renal tissue PLA2R analysis are sensitive and specific in detecting PMN. Baseline serum anti-PLA2R antibody and anti-PLA2R antibody quantification trends are related to prognosis of PMN. So they can be incorporated as additional biomarker.