Galectins are a family of beta-galactoside-binding proteins that are characterised by their carbohydrate recognition domain (CRD) and include galectin-1 and galectin-3. These galectins have been implicated in numerous diseases due to their pleiotropic nature, including cancer and fibrosis, with therapeutic inhibitors being clinically developed to block the CRD. One of the early methods developed to characterise these galectins was the hemagglutination of red blood cells. Although it is insightful, this approach has been hampered by a lack of sensitivity and accurate quantification of the agglutination observed. In this study, we aimed to validate a more precise and quantitative method to enable the further investigation of differences between galectins in respect to agglutination induction in different blood groups, as well as the characterisation of small molecule inhibitors. Quantification of hemagglutination was shown to be optimal using U-bottom plates imaged and analysed with FIJI ImageJ rather than flat-bottom plates read for absorbance on an optical density plate reader. Galectin-3-induced red blood cell agglutination efficacy increased significantly from blood group O to A to B. However, for both the galectin-1 monomer and concatemer, a more comparable effect was observed between blood group B and O, but with more potent effects than in blood group A. Inhibition assays for both galectin-3 and galectin-1 induced-hemagglutination were able to demonstrate clear concentration responses and expected selectivity profiles for a set of small-molecule glycomimetics, confirming the historical profiles obtained in biochemical binding and functional cellular assays.

OBJECTIVE: Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests.
METHODS: In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard.
RESULTS: Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ2 = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test.
CONCLUSIONS: There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability.

As an alternative to the classical method of erythrocyte hemagglutination, a latex agglutination assay based on the interaction of influenza viruses with the sialoglycoprotein fetuin immobilized on the surface of polystyrene microspheres has been developed. Twelve influenza A virus strains of different subtypes and two influenza B viruses of different lines were tested. Simultaneous titration of viruses using the classical hemagglutination test and the proposed latex agglutination assay showed similar sensitivity and a high degree of correlation (R = 0.94). The obtained microspheres can be used for titration of viruses that recognize and bind sialylated glycans as receptors. In particular, latex aggregation was also induced by the Newcastle disease virus.

Cystic echinococcosis (CE) is one of the neglected tropical diseases announced by the World Health Organization. In the period entered with the Coronavirus disease-2019 pandemic, the fight against such diseases has become even more difficult. In our study, we aimed to make inferences about the effects of the pandemic on the diagnosis of the disease by evaluating the number and results of CE indirect hemagglutination test (IHA) before and during the pandemic.
The number of IHA test requests and positivity rates in the 30-month periods before and after March 11, 2020, when the first case was seen in our country, were evaluated retrospectively. Statistical analysis was made with SPSS version 23 (SPSS, Chicago, IL, USA) program.
The results of 1444 patients before the pandemic and 870 patients during the pandemic period were examined. The difference between IHA positivity rates, which was found to be 18.49% before the pandemic and 14.6% during the pandemic, was statistically significant (p=0.016). The positivity rates of women and men were found to be statistically similar in both periods (pbefore=0.621, pafter=0.238). The age group with the highest IHA positivity rate was 20-39 in both periods, and the difference between the positivity rates of the age groups was statistically significant (p<0.001).
A significant decrease was observed in the rate of IHA positivity during the pandemic period. The status of no increase in positivity rates despite a significant decrease in IHA tests makes us think that the diagnosis may be missed in some patients or that there could be disruptions in their follow-up. For this reason, in order to continue the fight successfully against CE, which is an important public health problem for our country, early diagnosis and regular follow-ups should be emphasized with educations, and the laboratory-clinician communication should be strengthened in order to use tests more efficiently.
Kistik ekinokokkoz (KE) Dünya Sağlık Örgütü tarafından açıklanan ihmal edilmiş tropikal hastalıklardan biridir. Koronavirüs hastalığı-2019 pandemisiyle birlikte girilen süreçte bu tür hastalıklarla mücadele daha da zorlaşmıştır. Çalışmamızda pandemi öncesinde ve sürecinde KE indirekt hemaglütinasyon testi (IHA) sayılarını ve sonuçlarını değerlendirerek, pandeminin hastalık tanısı üzerindeki etkileri hakkında çıkarımlarda bulunmayı amaçladık.
Ülkemizde ilk olgunun görüldüğü 11 Mart 2020 tarihinden önceki ve sonraki 30 aylık süreçte IHA test istem sayıları ve pozitiflik oranları geriye dönük olarak değerlendirildi. İstatistiksel analizler SPSS versiyon 23 (SPSS, Chicago, IL, ABD) programı kullanılarak yapıldı.
Pandemi öncesinde 1444 ve pandemi döneminde 870 hastaya ait sonuç incelenmiştir. Pandemiden önce %18,49, pandemi sürecinde 14,6 olarak saptanan IHA pozitiflik oranları arasındaki fark istatistiksel olarak anlamlı bulunmuştur (p=0,016). Kadın ve erkeklerde pozitiflik oranları her iki dönemde de istatistiksel olarak benzer saptanmıştır (pönce=0,621, psonra =0,238). IHA pozitiflik oranının en fazla saptandığı yaş grubu her iki dönemde de 20-39 olup, yaş gruplarının pozitiflik oranları arasındaki fark istatistiksel olarak anlamlı bulunmuştur (p<0,001).
Pandemi döneminde IHA pozitiflik oranında anlamlı bir azalma görülmüştür. IHA test istem sayısında belirgin bir düşüş gözlenmesine rağmen, pozitiflik oranlarında artış saptanmaması; bazı hastalarda tanının atlanmış olabileceğini veya hastaların takiplerinde aksamalar yaşanmış olabileceğini düşündürmektedir. Sonuç olarak ülkemiz için önemli bir halk sağlığı problemi olan KE mücadelesine başarıyla devam edebilmek adına farkındalık eğitimleriyle birlikte erken tanı ve düzenli takiplerin önemi vurgulanmalı; laboratuvar-klinik hekimleri arası iletişim güçlendirilerek testlerin daha etkili kullanılması sağlanmalıdır.

The decline in the population of ring-necked pheasants (Phasianus colchicus) in northwestern Germany since 2007 raises questions about the underlying causes. We therefore studied the growth and immune status of ring-necked pheasant chicks dependent on different feed composition. Here, 490 ring-necked pheasant chicks were raised in five groups up to nine weeks. While control groups C1 and C2 received sufficient crude protein (28%) and energy (12.5 MJ/Kg feed) according to current standards, group C2 was treated with cyclosporine eight hours prior to phythemagglutination (PHA) testing, serving as a positive immune suppressed control. Group V1 was fed with reduced protein (20%) but optimal energy content (12.5 MJ/Kg feed), group V2 was fed with sufficient protein (28%) and reduced energy content (10 MJ/kg feed) whereas group V3 was fed reduced crude protein (20%) and reduced energy content (10MJ/kg feed). On all chicks, health status was checked each week, and 20 birds of each group were weighed randomly per week. PHA-testing was performed on 12 birds of each group to study the in vivo non-specific activation of lymphocytes at week 2, 4, 6, 7, 8 and 9. In addition, hemolysis-hemagglutination-assay (HHA) was performed on each of the PHA-tested chicks, which were subsequently euthanized and dissected. Histopathologic examinations of 5 birds that were randomly chosen were performed. The PHA-test results demonstrate significant differences between control (C1, C2) and experimental groups (V1-V3) in several developmental stages. According to the HHA results, weekly testing detected a significant increase of titres per week in all groups without significant differences. Here, only hemagglutination and no lysis of samples was observed. It seems appropriate to conclude that during their first weeks of life, protein content is of higher importance in ring-necked pheasant chicks than energy intake. In particular T-cell response is significantly reduced, which indicate a weaker immune system resulting in a higher risk for clinical diseases. Therefore, we assume that protein i.e. insect availability is a highly important co-factor in the free-ranging population dynamics, and is linked to declines of the northwestern German population.

OBJECTIVE: This study aims to evaluate the serological, radiological and epidemiological analysis of suspected cystic echinococcosis patients, and to assess the positivity rate in the region.
UNASSIGNED: The retrospective study was conducted at Bursa Uludag University Hospital, Turkey and comprised data from January 2009 to December 2017 related to patients of either gender with suspected cystic echinococcosis who underwent indirect haemagglutination testing. Demographic and clinical data of patients who tested positive were analysed. Statistical analysis was done using SPSS 23.
RESULTS: Of the 3910 patients with a mean age of 41.6±19.35 years (range: 0-93 years) who underwent indirect haemagglutination testing, 692(17.7%) tested positive; 390(56.4%) females, and 302(43.6%) males. The highest seropositivity rate 107(15.5%) was observed in 2011, followed by 104(15%) in 2016. Seropositive cases were predominantly seen in those aged 40-49 years 131 (18.9%), followed by those aged 50-59 years 124 (17.9%).
CONCLUSIONS: Cystic echinococcosis was found to be a public health problem in South Marmara region of Turkey.

Cystic echinococcosis (CE) is a parasitic disease that has been known for years in helminth diseases and it is important as human and animal health problem in many parts of the world and in our country due to economic losses. In this study, it was aimed to retrospectively evaluate the distribution of anti-E. granulosus-IgG antibodies in patients with pre-diagnosis of CE that referred to parasitology laboratory between January 2013-December 2018.
Commercial kit was used for indirect hemaglutination (IHA), indirect fluorescent antibody test (IFAT) and Western blot (WB) methods using sera from patient samples was applied according to the kit proposal. In addition, patient materials for CAM, CSF and blood for which polymerase chain reaction (PCR)/QPCR tests were requested were examined.
Sera of the patients who were tested with at least one of the IHA, IFAT and WB methods or a combination of these methods, and 443 cases out of 2.283 cases were found to be E. granulosus seropositive. It was determined that 369 (62.03%) of 443 positive patients were female and 330 (37.97%) were male patients. Among these patients, 87 patients whose IFAT and/or IHA tests were negative were found to have positive results with the WB method. IFAT or IHA test results of 13 patients with negative WB tests were found to be positive. Four patients were identified with both tests positive but WB test results negative. In addition, 36 of 72 patients who underwent PCR/QPCR tests were found to be positive.
As a result of a six-year retrospective screening, 22% of the cases were found to be positive, and it was concluded that the prevalence of CE is high and the use of a single test may be insufficient in the diagnosis of CE, therefore, test combinations will increase the sensitivity and reliability in reaching the correct diagnosis.
Kistik ekinokokkozis (KE), helmint hastalıkları içinde yıllardır bilinen, insan ve hayvan sağlığının yanı sıra sebep olduğu ekonomik kayıplar nedeniyle dünyanın birçok bölgesinde ve ülkemizde halk sağlığı problemi olarak önemini koruyan paraziter bir hastalıktır. Bu çalışmada Ocak 2013-Aralık 2018 tarihleri arasında parazitoloji laboratuvarına tetkik için başvuran ve ön tanısı KE olan hastaların anti-E. granulosus-IgG antikorlarının dağılımının retrospektif olarak değerlendirilmesi amaçlanmıştır.
İndirekt hemaglütinasyon (IHA), indirekt flerosan antikor testi (IFAT) ve Western blot (WB) yöntemlerinde ticari kit önerisine göre test prosedürü hasta numunelerinden elde edilen serumlar kullanılarak uygulanmıştır. Ayrıca, polimeraz zincir reaksiyon (PZR)/QPZR testleri yapılması istenilen kist aspirasyon mayii, beyin omurilik sıvısı ve kan hasta materyalleri incelenmiştir.
Serum örnekleri IHA, IFAT ve WB yöntemlerinin en az biri veya bu yöntemlerin kombinasyonları ile test edilerek, toplam 2283 olgudan 443 olgunun E. granulosus seropozitif olduğu belirlenmiştir. Dört yüz kırk üç pozitif hastanın 369’unun (%62,03) kadın ve 330’unun (%37,97) erkek hastalardan oluştuğu tespit edilmiştir. Bu hastalardan, IFAT ve/veya IHA testi negatif olan 87 hastanın WB yöntemi ile test sonucunun pozitif olduğu tespit edilmiştir. WB testi negatif olan 13 hastanın IFAT veya IHA test sonuçları ise pozitif olarak tespit edilmiştir. Her iki testin pozitif fakat WB test sonucunun ise negatif olduğu 4 hasta saptanmıştır. Ayrıca, PCR/QPCR testleri yapılan toplam 72 hastanın da 36’sının pozitif olduğu tespit edilmiştir.
Altı yıllık bir retrospektif tarama sonucunda olguların %22’sinin pozitif tespit edilerek KE yaygınlığının yüksek olduğu ve KE tanısında tek bir testin kullanımının yetersiz kalabileceği, bu nedenle test kombinasyonlarının doğru tanıya ulaşmada duyarlılığı ve güvenilirliği artıracağı kanaatine varılmıştır.

During the ongoing COVID-19 pandemic, serology has suffered several manufacturing and budget bottlenecks. Kode technology exposes exogenous antigens on the surface of cells; in the case of red blood cells, modified cells are called kodecytes, making antibody-antigen reactions detectable by the old-fashioned hemagglutination test. In this commentary, we review evidence supporting the utility of SARS-CoV-2 Spike kodecytes for clinical diagnostic purposes and serosurveys in resource-poor settings.

Automated chemiluminescent microparticle immunoassays (CMIAs) are the most common first step at high-volume laboratories for syphilis screening. If the initial screening test is reactive, 1 more treponemal test is required, resulting in increased cost. In this multicenter study, we aimed to determine the correlation between the CMIA signal-to-cutoff ratio (S/Co) and the confirmatory tests to reduce unnecessary confirmatory testing.
Eight hospitals from 5 provinces participated in this study. All laboratories used Architect Syphilis TP CMIA (Abbott Diagnostics, Abbott Park, IL) for initial screening. Treponema pallidum hemagglutination (TPHA), rapid plasma reagin (RPR), and fluorescent treponemal antibody absorption (FTA-ABS) were used as confirmatory tests according to the reverse or European Centre for Disease Prevention and Control algorithms. A receiver operating characteristic analysis was used to determine the optimal S/Co ratio to predict the confirmation results.
We evaluated 129,346 serum samples screened by CMIA between January 2018 and December 2020. A total of 2468 samples were reactive; 2247 (91%) of them were confirmed to be positive and 221 (9%) were negative. Of the 2468 reactive specimens, 1747 (70.8%) had an S/Co ratio ≥10.4. When the S/Co ratios were ≥7.2 and ≥10.4, the specificity values were determined to be 95% and 100%, respectively. In a subgroup of 75 CMIA-positive patients, FTA-ABS was performed and 62 were positive. Among these FTA-ABS-positive patients, 24 had an S/Co ratio <10.4, and negative TPHA and RPR.
We propose a potentially cost-effective reverse screening algorithm with a treponemal CMIA S/Co ratio ≥10.4, obviating the need for secondary treponemal testing in about 71% of the screening-reactive samples. This would substantially reduce the confirmatory testing volume and laboratory expenses. However, in high-risk group patients with CMIA positive results, S/Co ratio <10.4, and negative TPHA and RPR, FTA-ABS may be used for confirmation.

Rabies vaccine preparations are quantitatively assayed for potency using the in-vivo challenge National Institute of Health (NIH), the main test that consumes a high number of animals, takes a long time, and has wide variability. The Rapid focus fluorescent inhibition (RFFIT) and the passive hemagglutination (PHA) tests, the two serologically based tests, were also used for such purpose. In this study, we aimed to evaluate and correlate the potency of the NIH, RFFIT, and PHA tests according to the World Health Organization (WHO) validity criteria, aiming to validate the use of RFFIT or PHA test as a substitute to the NIH test for determining the potency of commercially available Rabies vaccine preparations. The results showed that, the three tests can be successfully used; however, a higher correlation between RFFIT and NIH than PHA and NIH was recorded (Pearson correlation = 1). The potency of rabies vaccine preparations using NIH, RFFIT, and PHA were 3.73, 3.51, and 4.50, respectively. NIH is the main test for the determination of vaccine potency carried out by conducting 25 experiments and consuming about 5,000 mice compared to 1,200 mice used with RFFIT and 1,000 mice used with PHA test. Taken together, we concluded that (i) in some tested preparations, both RFFIT and PHA tests gave comparable results, and they can be used interchangeably; (ii) RFFIT could successfully replace NIH test, but not PHA; (iii) RFFIT and PHA tests are faster, more accurate, more economic, and more sensitive than NIH; nevertheless, PHA needs further investigations; and (iv) both RFFIT and NIH tests complement and reinforce each other as they provide a comprehensive picture of the product potency.