scholarly journals Biochip Technology for the Serological Diagnosis of Bullous Pemphigoid

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
Haik Zarian ◽  
Andrea Saponeri ◽  
Anna Michelotto ◽  
Edoardo Zattra ◽  
Anna Belloni-Fortina ◽  
...  
Keyword(s):  
Bullous Pemphigoid ◽  
Indirect Immunofluorescence ◽  
Immunofluorescence Assay ◽  
Dermoepidermal Junction ◽  
Suitable Alternative ◽  
Clinical Criteria ◽  
Specific Proteins ◽  
Laboratory Investigations ◽  
Circulating Autoantibodies

Bullous pemphigoid is an autoimmune blistering skin disease characterized by the presence of circulating autoantibodies which recognize specific proteins of the epidermis and dermoepidermal junction. Diagnosis is based on clinical criteria and laboratory investigations, notably histology, direct and indirect immunofluorescence, and ELISA. This study describes a new immunofluorescence assay for parallel determination of anti-BP180 and anti-BP230 based on recombinant antigenic substrates. The aim of the study was to detect BP180 and BP230 autoantibodies by BIOCHIP technology using both a specially designed recombinant BP180-NC16A protein and cells expressing the BP230-gc antigen fragment. 18 patients with bullous pemphigoid were included in the study. Autoantibodies to BP180 were detected by the BIOCHIP technique in 83.33% of patients with clinical, serological, and immunohistological confirmed bullous pemphigoid while autoantibodies against BP230-gC were detected only in 39% of patients. The detection of anti-BP180-NC16A and anti-BP230-gC by a new biochip-based immunoassay is a suitable alternative to indirect immunofluorescence and ELISA. This method has the advantage of easily discriminating the different autoantibody specificities. The BIOCHIP method is faster, cheaper, and easy to use when compared with the ELISA approach. For this reason, the new method could be used as an initial screening test to identify patients with bullous pemphigoid, and doubtful results could then be confirmed by ELISA.

scholarly journals Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Christiane Radzimski ◽  
Christian Probst ◽  
Bianca Teegen ◽  
Kristin Rentzsch ◽  
Inga Madeleine Blöcker ◽  
...  
Keyword(s):  
Autoimmune Hepatitis ◽  
Indirect Immunofluorescence ◽  
Rat Kidney ◽  
Immunofluorescence Assay ◽  
Hek293 Cells ◽  
Indirect Immunofluorescence Assay ◽  
Biliary Cirrhosis ◽  
Dependent Manner ◽  
Hyperimmune Serum

Autoantibodies against soluble liver antigen (SLA) are specific markers for autoimmune hepatitis (AIH) type 1. In contrast to the determination of other AIH-associated autoantibodies by indirect immunofluorescence assay (IFA), detection of anti-SLA relied up to now on ELISA or immunoblot based on bacterially expressed recombinant protein. In order to develop a complementary IFA substrate, SLA isoform 1 was recombinantly produced in the human cell line HEK293 and controlled by a rabbit hyperimmune serum against SLA. The recombinant cells were used in IFA (RC-IFA) to analyze sera from 20 AIH patients with anti-SLA positivity predetermined by ELISA together with 80 controls (20 anti-SLA negative AIH, 15 primary biliary cirrhosis, 15 HCV, and 30 healthy blood donors). Using RC-IFA, anti-SLA was detected in all ELISA positive AIH sera but in none of the controls. Furthermore, a cytosolic fraction of HEK293 containing SLA was able to neutralize the autoantibodies in all positive sera in a dose-dependent manner. HEK293 cells expressing SLA are a valid substrate for the serodiagnosis of AIH relevant autoantibodies by IFA. In concert with cryosections of primate liver, rat kidney, rat liver, rat stomach, and HEp-2 cells, they enable the parallel determination of all autoantibodies associated with autoimmune liver diseases.

Transmission experiments of cilia-associated respiratory bacillus in mice, rabbits and guineapigs

1989 ◽  
Vol 23 (2) ◽  
pp. 96-102 ◽  
Author(s):  
S. Matsushita ◽  
H. Joshima ◽  
T. Matsumoto ◽  
K. Fukutsu
Keyword(s):  
Indirect Immunofluorescence ◽  
Direct Contact ◽  
Immunofluorescence Assay ◽  
The Other ◽  
Indirect Immunofluorescence Assay ◽  
Primary Method ◽  
Airborne Transmission ◽  
Histological Changes ◽  
Observation Period

Transmission experiments of cilia-associated respiratory (CAR) bacillus were performed in mice in order to clarify the principal route of the infection, and in rabbits and guineapigs in order to examine their susceptibility. Determination of the infection was evaluated serologically by the indirect immunofluorescence assay (IFA) technique and histologically by the presence of CAR bacillus in the airways. BALB/c mice were intranasally inoculated with the SMR strain of CAR bacillus. The IFA antibody to the bacteria in these mice rose to more than 1 : 160 at 4 weeks postinoculation (PI) and the mice were utilized as transmitters for the following experiments. One out of 15 uninfected mice kept in intracage contact with infected mice became infected from 4 weeks after contact. Incidence of contact infection increased thereafter. On the other hand, there was no evidence of infection in the uninfected mice housed in the separate cages from the cage in which infected mice were housed throughout the 12-week observation period. The primary method of CAR bacillus transmission seems to be direct contact with infected mice or fomites contaminated by infected mice; airborne transmission appears to be of little importance. Rabbits and guineapigs were also intranasally inoculated with the SMR strain of CAR bacillus. IFA antibodies were positively detected by 4 weeks PI, but no CAR bacillus nor histological changes relating to the infection were observed in the airways of either species. It is suggested that rat origin CAR bacillus can transmit to rabbits and guineapigs, and that the infection can spread to other species of rodents and rabbits.

scholarly journals New-Onset Bullous Pemphigoid in a COVID-19 Patient

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Natalie Olson ◽  
David Eckhardt ◽  
Angela Delano
Keyword(s):  
Bullous Pemphigoid ◽  
Type Ii Diabetes Mellitus ◽  
Punch Biopsy ◽  
The Body ◽  
Direct Immunofluorescence ◽  
Class Iii ◽  
Dermoepidermal Junction ◽  
Clinical Criteria ◽  
History Of ◽  
Important Addition

This manuscript presents a report of bullous pemphigoid rash associated with COVID-19 for the first time. The objective of this manuscript is to present a unique dermatological case in the setting of a COVID-19-positive infection to further recognize the virus symptomatology. A 37-year-old female with a past medical history of class III obesity, type II diabetes mellitus, and hypertension presented to the emergency department in September 2020 with inpatient and outpatient follow-up through to November 2020. The patient denied any personal or family history of skin disorders. The patient tested positive for COVID-19 prior to hospitalization and presented to the hospital with severe, persistent, pruritic rash meeting dermatopathological, serologic, and clinical criteria for bullous pemphigoid diagnosis. Histopathology H&E punch biopsy from her left flexor wrist demonstrated epidermal keratinocyte necrosis, subepidermal vesiculation with eosinophils, gossamer stranding of the papillary dermis, and subepidermal edema. Direct immunofluorescence punch biopsy from her left flexor wrist demonstrated strong linear IgG staining at the dermoepidermal junction, with weaker and focal linear C3 staining. Antigen-specific serology was consistent with bullous pemphigoid. There was no previously reported cutaneous association of COVID-19 infection with bullous pemphigoid making this case an important addition to the body of evidence helping to identify bullous pemphigoid in the setting of viral infection.

Determination of the Viability of Toxoplasma gondii in Cured Ham Using Bioassay: Influence of Technological Processing and Food Safety Implications

2010 ◽  
Vol 73 (12) ◽  
pp. 2239-2243 ◽  
Author(s):  
SUSANA BAYARRI ◽  
MARÍA J. GRACIA ◽  
REGINA LÁZARO ◽  
CONSUELO PÉREZ-ARQUILLUÉ ◽  
MONTSERRAT BARBERÁN ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Horizontal Transmission ◽  
Meat Product ◽  
Immunofluorescence Assay ◽  
Nitrite Concentration ◽  
Bioassay Technique ◽  
Technological Processing ◽  
Undercooked Meat ◽  
Porcine Tissues

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii and distributed worldwide. Ingestion of viable cysts from infected raw or undercooked meat is an important route of horizontal transmission of the parasite to humans. Little information is available concerning the effect of commercial curing on cysts of T. gondii. This study is the first in which the influence of processing of cured ham on the viability of T. gondii has been evaluated, using bioassay to assess the risk of infection from eating this meat product. Naturally infected pigs were selected for the study, and a mouse concentration bioassay technique was used to demonstrate viable bradyzoites of T. gondii in porcine tissues and hams. No viable parasites were found in the final product (14 months of curing) based on results of the indirect immunofluorescence assay and histological and PCR analyses. Our results indicate that the consumption of hams cured as described here poses an insignificant risk of acquiring toxoplasmosis. However, additional studies are required to evaluate the safety of ham products cured under different conditions of curing time, salt, and nitrite concentration.

scholarly journals The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anne E. Tebo ◽  
Robert L. Schmidt ◽  
Kamran Kadkhoda ◽  
Lisa K. Peterson ◽  
Edward K. L. Chan ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
Nuclear Staining ◽  
International Consensus ◽  
In Vitro Diagnostic ◽  
Traditional Pattern

Abstract Background To evaluate the interpretation and reporting of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) using HEp-2 substrates based on common practice and guidance by the International Consensus on ANA patterns (ICAP). Method Participants included two groups [16 clinical laboratories (CL) and 8 in vitro diagnostic manufacturers (IVD)] recruited via an email sent to the Association of Medical Laboratory Immunologists (AMLI) membership. Twelve (n = 12) pre-qualified specimens were distributed to participants for testing, interpretation and reporting HEp-2 IFA. Results obtained were analyzed for accuracy with the intended and consensus response for three main categorical patterns (nuclear, cytoplasmic and mitotic), common patterns and ICAP report nomenclatures. The distributions of antibody titers of specimens were also compared. Results Laboratories differed in the categorical patterns reported; 8 reporting all patterns, 3 reporting only nuclear patterns and 5 reporting nuclear patterns with various combinations of other patterns. For all participants, accuracy with the intended response for the categorical nuclear pattern was excellent at 99% [95% confidence interval (CI): 97–100%] compared to 78% [95% CI 67–88%] for the cytoplasmic, and 93% [95% CI 86%–100%] for mitotic patterns. The accuracy was 13% greater for the common nomenclature [87%, 95% CI 82–90%] compared to the ICAP nomenclature [74%, 95% CI 68–79%] for all participants. Participants reporting all three main categories demonstrated better performances compared to those reporting 2 or less categorical patterns. The average accuracies varied between participant groups, however, with the lowest and most variable performances for cytoplasmic pattern specimens. The reported titers for all specimens varied, with the least variability for nuclear patterns and most titer variability associated with cytoplasmic patterns. Conclusions Our study demonstrated significant accuracy for all participants in identifying the categorical nuclear staining as well as traditional pattern assignments for nuclear patterns. However, there was less consistency in reporting cytoplasmic and mitotic patterns, with implications for assigning competencies and training for clinical laboratory personnel.

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