Human Neutrophil Alloantigen-3a Alloimmunization among Pregnant Korean Women / 대한수혈학회지

BACKGROUND: Alloantibodies against human neutrophil alloantigen (HNA)-3a are associated with severe and fatal transfusion related acute lung injury (TRALI). HNA-3 genotyping and HNA-3a antibody (Ab) identification are essential to diagnosis and prevention of TRALI caused by HNA-3a Ab. However there had been no laboratory for HNA-3a Ab identification in Korea. The aims of this study were to establish the HNA-3a Ab test in Korea and to estimate the incidence of HNA-3a alloimmunization among pregnant Korean women. METHODS: HNA-3a homozygotes and HNA-3b homozygotes were identified by HNA-3 genotyping. Three HNA-3a homozygotes and three HNA-3b homozygotes are included in the granulocytes panel, which consisted of 10 donors for granulocytes. Sera from 650 pregnant Korean women were tested for granulocyte Ab using a mixed passive hemagglutination assay (MPHA). When a HNA-3a Ab was detected, the woman's HNA-3 was typed to support her HNA-3a alloimmunization. RESULTS: MPHA showed positive reactions in the sera from 26 women (4.0%, 26/650). HLA Abs were detected in 18 women (2.8%, 18/650), among whom HNA Abs were identified simultaneously in 7 women. Granulocyte Abs were detected in sera from 15 women (2.3%, 15/650). The incidence of HNA-3a, HNA-1b, HNA-1a, HNA-2a, and unidentified HNA Abs among pregnant Korean women was 0.77% (5/650), 0.77% (5/650), 0.62% (4/650), 0.15 (1/650), and 0.31% (2/650), respectively. CONCLUSION: In this study, we established the HNA-3a Ab test using MPHA for diagnosis and prevention of TRALI caused by HNA-3a Ab. The incidence of HNA-3a Ab in pregnant Korean women was 0.77% (5/650).

Ficus religiosa L.: Callus, Suspension Culture and Lectin Activity in Fruits and In vitro Regenerated Tissues.

Ficus religiosa L. is a tree of immense cultural heritage in Asian countries. It is respected by followers of many religions and faiths. Fruits of Ficus religiosa L. are the ‘figs’ and possess many medicinal properties reported in ethnomedicinal and pharmacological studies. These medicinal properties range from antidiabetic, anticancer, anticonvulsant, antimicrobial, antiviral and antioxidant activities. In the figs, pollination takes place with ‘wasp’. Till date no work on ‘fruit tissue culture’ has been reported in this species. For the first time the callus cultures have been developed using ‘fig fruits’. Fruit callus was multiplied on solid medium using 2 mm to 3 mm diameter fruits. During the present study, lectin/hemagglutinin activity was detected in fruits and fruit callus extracts for the first time. Both In vivo fruits (figs) and In vitro fig callus were used to assay the hemagglutinin activity using pronase treated and untreated rabbit blood erythrocytes. Fruit extract showed 4-8 times more hemagglutination activity in presence of pronase treated erythrocytes. It is the first report of callus/suspension culture and detection of thermostable (up to 70ºC) hemagglutinin/lectin from fruits in this species. Preliminary biochemical characterization of the lectin activity e.g. metal ion requirement, EDTA, pH and temperature stability was carried out during the present study.

Bacteriostatic activity of con a lectin from Canavalia ensiformis.

The aim of this work was to explore the therapeutic applications of Con A lectin from Canavalia ensiformis and to explore its antibacterial activity. Activity of lectin was quantified by their ability to agglutinate erythrocytes using Hemagglutination assay. Characterization and purity of Con A lectin was evaluated by using SDS-PAGE analysis. The reversal of hemagglutination activity of lectin was evaluated by using the sugars namely; mannose, galactose, lactose, fructose, glucose. The antibacterial activity of lectins was tested against Streptococcus mutans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli using pour plate method. Amoxycillin was used as standard. At 250mg/ml concentration Con A lectin showed good bacteriostatic activity.

Evaluation on application of common diagnosis methods for schistosomiasis japonica in endemic areas of China Ⅱ Quantitive analysis of detection results for S.japonicum infection by IHA screening method and Kato-Katz technique / 中国血吸虫病防治杂志

Objectives To explore the correlation between the population characteristics of sero-reactivity with quantitative antibody based-IHA and the transmission parameters,such as epidemic situation,transmission status or infection trend in population.Methods The residents in one endemic administrative village were simultaneously examined by Kato-Katz technique for parasitological stool examination,as well as by immuno-diagnostic technique IHA for detection of IgG antibody against soluble egg antigen for two consecutive years.The results of examination were analyzed and compared on the diagnostic parameters of IHA,the correlation of the changes of positive rates and antibody levels of IHA with the changes of infection trend in population and the distribution of antibody levels between 'the true negative' and 'the true positive'.Results When Kato-Katz technique based on 2 stool samples,each read in 3 thick smears,was used as the reference,the overall sensitivity of IHA was high (from 77.27% to 85.48%) with a relatively poor specificity of lower than 60%,the negative predict value (NPV) was excellent of higher than 94%.The specificity of IHA decreased with the increase of the age in different age-groups of population,showing the highest among the younger less than 15 years old.The distribution trends of positive rates of antibody in different age groups by IHA showed similar to that of egg positive rate detected by Kato-Katz although the positive rates of IHA were higher than these by Kato-Katz,which showed that a higher false positive (from 41.90% to 44.56%) and a certain false negative (from 14.52% to 22.73%)existed in IHA.The positive rate of antibody decreased slowly among the individuals with S.japonicum infection,who received treatment.There was an overlap in the distribution of antibody levels between ' the true negative' and ' the true positive'.Conclusions Under the current schistosomiasis epidemic situation in China,IHA is valuable in the epidemiologic surveys.It should be of further deliberation applying IHA as the screening approaches for identification of target individuals for treatment or determination of the infection rate in community and IHA needs to be combined with the parasitological examination.

Comparison of Nine Different Qualitative HBsAg Assay Kits / 대한진단검사의학회지

BACKGROUND: Qualitative hepatitis B surface antigen (HBsAg) assay kits are still commonly used in Korea where hepatitis B virus (HBV) infection is endemic. The accurate determination of HBsAg plays a crucial role in the diagnosis and prevention of HBV infection, especially in endemic areas. The aim of this study was to compare the detection sensitivities of 9 qualitative HBsAg assay kits. METHODS: Seven pooled sera with HBsAg concentration ranging from 0.14 IU/mL to 29.96 IU/mL were prepared. The HBsAg concentration of each pooled serum was determined by a quantitative HBsAg assay, Architect HBsAg (Abbott Laboratories, Ireland). The fully automated immunoassay kits included Elecsys HBsAg (Roche Diagnostics, Germany) and Immulite 2000 HBsAg (DPC, USA) and the rapid tests included 5 immunochromatographic assay (ICA) kits and 2 reverse passive hemagglutination assay (RPHA) kits. RESULTS: Elecsys HBsAg (Roche Diagnostics) showed positive result in pooled serum with HBsAg concentration of 0.14 IU/mL, but Immulite 2000 HBsAg (DPC) showed negative result in the same concentration. Although ICA kits showed variable results among different assay kits, all of them showed negative results in pooled sera with HBsAg concentration of < or =1.89 IU/mL. Two RPHA kits showed negative results in pooled sera with HBsAg concentration of < or =7.98 IU/mL. CONCLUSIONS: Although ICAs were more sensitive than RPHAs, they had variable sensitivities for HBsAg and were less sensitive than the automated immunoassay kits. Therefore, ICAs and RPHAs should be used with caution in the screening tests for HBsAg and their sensitivities need to be improved.

Establishment of Mixed Passive Hemagglutination Assay Using In-House Anti-Rh(D) Sensitized Human O RBCs for the Detection of Granulocyte Specific Antibodies / 대한수혈학회지

BACKGROUND: Granulocyte specific antibodies are associated with several clinical conditions including febrile transfusion reaction and transfusion-related acute lung injury as well as immune neutropenias. The identification of granulocyte specific antibodies is important for the diagnosis of these disorders. However, there have been rarely confirmed clinical reports in Korea since the testing techniques are complicated and difficult to maintain. In this study, development of in-house indicator cells and renewedly establishment of the mixed passive hemagglutination assay (MPHA) as a serologic test to detect and identify granulocyte specific antibodies were conducted. METHODS: The in-house indicator cells for MPHA were made by sensitizing human Rh(D) positive O RBCs with human IgG anti-Rh(D) (DiaMed AG, Switzerland) and then combining with AHG anti-IgG (Immucor Inc., USA). To determine the optimal conditions, various combinations of anti-Rh(D) IgG sensitization strengths of indicator cells, microwell coated antigens (intact granulocyte vs. extracted granulocyte) and reaction conditions were compared. RESULTS: The best test conditions for MPHA were as follows: optimal results were obtained with the anti-Rh(D) sensitization dilutions of 1/64-1/192 and the reaction condition of 4 hours incubation at room temperature in humid chamber. Extracted granulocytes coated at the plate showed better results than intact granulocytes. HLA antigens were completely removed from extracted granulocyte antigens after acidified chloroquine treatment. CONCLUSION: Granulocyte MPHA using in-house anti-Rh(D) sensitized indicator cells was developed for the first time in Korea. The newly established MPHA would be effectively used for the diagnosis and treatment of disorders associated with granulocyte specific antigen-antibody reactions in Korea.

Comparison of Passive Hemagglutination and Indirect Immunofluorescence Assay for the Diagnosis of Scrub typhus / 감염과화학요법

The gold standard for the serologic detection of scrub typhus is the indirect immunofluorescence assay (IFA), however the passive hemagglutination assay (PHA) is commonly used method in Korea because of its availability, easy handling. We compared the sensitivity of PHA and IFA to compare their effectiveness in diagnosing scrub typhus. Out of 35 patients who were eventually diagnosed of scrub typhus by positive IFA, 19 patients (54%) were also tested positive for PHA during the acute phase (titer=1:80). Whereas 30 patients (85%) were positive in IFA during the acute phase. During the recovery phase 30 (85%) patients were positive in PHA, whereas all patients were found to be positive in IFA. Patients with 4-fold or greater rise of antibody were 12 (34.3%) in PHA, and 19 (54.3%) in IFA. We note that IFA has better sensitivity over PHA in detecting scrub typhus.

Comparison of Passive Hemagglutination and Indirect Immunofluorescence Assay for the Diagnosis of Scrub typhus / 감염과화학요법

The gold standard for the serologic detection of scrub typhus is the indirect immunofluorescence assay (IFA), however the passive hemagglutination assay (PHA) is commonly used method in Korea because of its availability, easy handling. We compared the sensitivity of PHA and IFA to compare their effectiveness in diagnosing scrub typhus. Out of 35 patients who were eventually diagnosed of scrub typhus by positive IFA, 19 patients (54%) were also tested positive for PHA during the acute phase (titer=1:80). Whereas 30 patients (85%) were positive in IFA during the acute phase. During the recovery phase 30 (85%) patients were positive in PHA, whereas all patients were found to be positive in IFA. Patients with 4-fold or greater rise of antibody were 12 (34.3%) in PHA, and 19 (54.3%) in IFA. We note that IFA has better sensitivity over PHA in detecting scrub typhus.