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Hepatitis B virus surface antigen in glomerular immune complex deposits of patients with systemic lupus erythematosus

Looi LM, Prathap K

Histopathology, 1982 Mar;6(2):141-7.
PMID: 7042512

In view of a high prevalence of hepatitis B virus infection in the Malaysian population, indirect immunofluorescence examination for hepatitis B surface antigen (HBsAg) was routinely performed on renal biopsy specimen at the University Hospital, Kuala Lumpur, over a 3-year period. Examination of renal tissue from 259 patients, including 47 with systemic lupus erythematosus (SLE), revealed 43 cases with HBsAg in glomerular immune complexes. A significantly high proportion (30/43) of these were SLE patients. The deposits were granular in nature, situated in both the capillary walls and mesangium and associated with immunoglobulin deposition. Morphological patterns of lupus nephritis involved were focal proliferative (one case), diffuse proliferative (23 cases) and membranous (six cases). None of these patients showed clinical evidence of liver disease. The significance of these findings remains uncertain, but the possibility exists that the hepatitis B virus may have a role in the pathogenesis of SLE in the tropics where both SLE and HBs antigenaemia are common.

Matched MeSH terms: Fluorescent Antibody Technique
Visualization of Bacteria-Mediated Gene Delivery Using High-Resolution Electron and Confocal Microscopy

Osahor AN, Ng AWR, Narayanan K

Methods Mol Biol, 2021;2211:29-40.
PMID: 33336268 DOI: 10.1007/978-1-0716-0943-9_3

Visual analysis of the gene delivery process when using invasive bacteria as a vector has been conventionally performed using standard light and fluorescence microscopy. These microscopes can provide basic information on the invasiveness of the bacterial vector including the ability of the vector to successfully adhere to the cell membrane. Standard microscopy techniques however fall short when finer details including membrane attachment as well as internalization into the cytoplasm are desired. High-resolution visual analysis of bacteria-mediated gene delivery can allow accurate measurement of the adherence and internalization capabilities of engineered vectors. Here, we describe the use of scanning electron microscopy (SEM) to directly quantify vectors when they are external to the cell wall, and confocal microscopy to evaluate the vectors when they have internalized into the cytoplasm. By performing the invasion procedure on microscope coverslips, cells can be easily prepared for analysis using electron or confocal microscopes. Imaging the invasion complexes in high resolution can provide important insights into the behavior of bacterial vectors including E. coli, Listeria, and Salmonella when invading their target cells to deliver DNA and other molecules.

Matched MeSH terms: Fluorescent Antibody Technique
Mesangial IGM in minimal change glomerular disease: a clinicopathological study in a Malaysian population

Looi LM, Wang F, Lam KL, Chua CT

Pathology, 1985 Jan;17(1):41-4.
PMID: 3889788

During a 6 yr period, 105 (69%) of 153 patients in whom a histological diagnosis of minimal change glomerular disease was made had renal biopsy tissue suitable for immunofluorescence examination. Thirty seven (35%) patients showed glomerular mesangial deposits of IgM. All the patients presented with the nephrotic syndrome. We found no significant difference in age and sex prevalence, presentation, response to therapy and glomerular morphology between IgM positive and IgM negative groups. This study suggests that there is no necessity to categorize IgM positive minimal change glomerular disease as a separate entity.

Matched MeSH terms: Fluorescent Antibody Technique
Phenotypic and microRNA characterisation of the neglected CD24+ cell population in MCF-7 breast cancer 3D spheroid culture

Boo L, Yeap SK, Ali NM, Ho WY, Ky H, Satharasinghe DA, et al.

J Chin Med Assoc, 2019 Nov 15.
PMID: 31770189 DOI: 10.1097/JCMA.0000000000000226

BACKGROUND: In vitro 3-dimensional spheroid culture has been widely used as model to enrich CD44CD24 cancer stem cells (CSC) with high ALDH1 activity. Although CD24subpopulation was known to be present in 3D spheroids and may influence cancer drug therapies, its characteristics and CSC properties were not well defined.
METHODS: In this study, CD24 population from the MCF-7 spheroid was sorted and subjected to spheroid formation test, stem cell markers immunofluorescence, invasion and migration test as well as microRNA expression profiling.
RESULTS: Sorted MCF-7 CD24 cells from primary spheroids were able to reform its 3D spheroid shape after 7 days in non-adherent culture conditions. In contrast to the primary spheroids, the expression of SOX-2, CD44, CD49f and Nanog were dim in MCF-7 CD24+ cells. Remarkably, MCF-7 CD24 cells were found to show high expression of ALDH1 protein which may have resulted in these cells exhibiting higher resistance against doxorubicin and cisplatin when compared to that of the parental cells. Moreover, microRNA profiling has shown that the absence of cancer stem cell properties were consistent with the downregulation of major cancer stem cells related pathways including Hedgehog, Wnt and MAPK signalling pathways. However, the upregulated pathways such as adherans junctions, focal adhesion and tight junction suggest that CD24+ cells were probably at an epithelial-like state of cell transition.
CONCLUSION: In conclusion, neglected CD24+ cells in MCF-7 spheroid did not exhibit typical breast CSCs properties. The presence of miRNAs and their analysed pathways suggested that these cells could be a distinct intermediate cell state in breast CSCs.

Matched MeSH terms: Fluorescent Antibody Technique
Seroepidemiology of malaria in Peninsular Malaysia: Plasmodium falciparum antibody profile of adults in four states

Thomas V, Bin HK, Leng YP

Trans R Soc Trop Med Hyg, 1980;74(3):375-80.
PMID: 7001690

In 1973, 2610 sera were collected from adults living in 22 localities in four states in Peninsular Malaysia and tested by IFAT for Plasmodium falciparum antibodies. A larger number of thin films were examined. The attack phase of the Malaria Eradication Programme (MEP) in these areas was started between 1968 and 1973. The results showed that the highest prevalence rates and geometrical mean reciprocal titres (CMRT) were among adults from Kelantan where the antibody prevalence varied greatly among the adults and there was active transmission in at least three areas. The values were lowest for Kedah. The P. falciparum antibody prevalence rates were higher than the parasite rates as revealed in single thin film examinations but a number of the positive sera were reactive only at low titres. The low concentration probably indicated the residual antibody from cured cases or past infections and cross reactions to P. vivax and P. malariae infections. The strong reactions probably indicated current P. falciparum transmission as shown by positive thin films. The present study showed that the antibody profile of adults, as shown by IFAT, is of considerable value in assessing the malaria situation in a given area and that it would be useful as a malariometric tool in epidemiological studies to evaluate the progress of malaria eradication/control programmes.

Matched MeSH terms: Fluorescent Antibody Technique
Mammals and scrub typhus ecology in peninsular Malaysia

Muul I, Liat LB, Walker JS

Trans R Soc Trop Med Hyg, 1975;69(1):121-30.
PMID: 806995

The overall comparisons of habitats are given in (Table III). The habitats are arranged in order of extent of alterations by man, with the least disturbed at the top. The highest average blood isolation rates came from the least disturbed areas. The highest monthly maximal rickettsial isolation rates from blood and maximal prevalence rates of antibody per month were also obtained at Bukit Lanjan, the habitat least altered by activities of man. The lowest average blood isolation rate (6%) and the lowest monthly maximal rickettsial isolation and antibody prevalence rates were obtained at Bukit Mandol, the habitat most extensively and intensively altered by man. The intermediate habitats had intermediate rates. We caution anyone interpreting these observations, however, in terms of human disease, which seem to be associated with hyperendemic foci. Here we are not dealing with hyperendemicity from the standpoint of human disease, but present evidence of widespread endemicity from which hyperendemic foci may derive. Also, we have not yet identified the prevalent strains and do not know their infectivity to man.

Matched MeSH terms: Fluorescent Antibody Technique
Fulltext Immunofluorescence (IF) pattern, autoantigens of anti-neutrophil cytoplasmic antibodies (ANCA) and their clinical associations

Masita Arip, Marlyn Mohammad, Salbiah Nawi, Shahnaz Murad

Jurnal Sains Kesihatan Malaysia, 2010;8(2):37-41.
MyJurnal

Antineutrophil cytoplasmic antibodies (ANCA) are autoantibodies directed against primary granules of neutrophils and monocytes' lysosomes. In general, c-ANCA is strongly associated with vasculitic disorders mainly in ANCA-associated systemic vasculitis (AASV). p-ANCA have been identified in several diseases such as primary (AASV) and secondary vasculitis such as collagen vascular diseases, rheumatoid arthritis and inflammatory bowel diseases given the term 'ANCA-associated disease.' The objective of this study was to determine the rate of ANCA positivity by indirect immunofluorescent (IF) and enzyme linked immunosorbent assay (ELISA) and its association with AASV and ANCA associated diseases. Serum from patients with history suspicion of systemic vasculitis were tested for ANCA by IF. Those samples positive for ANCA by IF were further tested for antibodies against myeloperoxidase (MPO) and proteinase 3 (PR3) using the ELISA. Clinical data from medical records were obtained and analyzed. Of 468 samples, a total of 110 were positive for ANCA by IF. IF results showed a p-ANCA pattern in 96 patients and c-ANCA in 14. Of 110 IF positive ANCA, 45 patients were positive by ELISA. Seventeen were positive for MPO-ANCA, 9 were PR3-ANCA positive and 19 were both MPO and PR3 positive. Only 2 patients were classified AASV ie Wegener granulomatosis and the other with microscopic polyangiitis. The remaining patients (n = 108) may be classified as ANCA associated diseases. Our study showed that pANCA (87.3%) was the more common ANCA pattern and 40.9% of IF positive samples were positive for PR3- and MPOANCA.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Ghrelin stimulates growth hormone release from the pituitary via hypothalamic growth hormone-releasing hormone neurons in the cichlid, Oreochromis niloticus

Ogawa S, Liu X, Shepherd BS, Parhar IS

Cell Tissue Res, 2018 Nov;374(2):349-365.
PMID: 29934855 DOI: 10.1007/s00441-018-2870-6

Ghrelin, a gut-brain peptide hormone, is implicated in a multiplicity of biological functions, including energy homeostasis and reproduction. Neuronal systems that are involved in energy homeostasis as well as reproduction traverse the hypothalamus; however, the mechanism by which they control energy homeostasis is not fully understood. The present study analyzes the anatomical relationship of neurons expressing gonadotropin-releasing hormone (GnRH), neuropeptide Y (NPY) and growth hormone-releasing hormone (GHRH) in a cichlid, tilapia (Oreochromis niloticus). Additionally, we examine in vivo effects of ghrelin on these hypothalamic neurons and plasma growth hormone (GH) and insulin-like growth factor-1 (IGF-1) levels. Double-immunofluorescence showed neuronal fiber associations between GnRH, NPY and GHRH in the brain and pituitary. Intracerebroventricular injection of ghrelin had no effect on numbers, soma size, or optical density of GnRH and NPY neurons, whereas the number of GHRH neurons was significantly decreased in the animals injected with ghrelin when compared to controls, which may indicate administered ghrelin promoted GHRH release. Plasma GH and pituitary GH mRNA levels were significantly increased in the animals injected with ghrelin. These results suggest that central administration of ghrelin primarily act on hypothalamic GHRH neurons to stimulate GH release from the pituitary in the tilapia.

Matched MeSH terms: Fluorescent Antibody Technique
Fulltext In vivo response of GsdmA3(Dfl)/+ mice to topically applied fish oil - effects on cellular markers and macrophages

Zulfakar MH, Porter RM, Heard CM

FEBS Open Bio, 2016 08;6(8):827-34.
PMID: 27516961 DOI: 10.1002/2211-5463.12095

Psoriasis is an incurable autoimmune disease characterized by patches of abnormal red, itchy and scaly skin. This work examined the modulation of inflammation, hyperproliferation and immune cell markers following topical application of fish oil (FO) in comparison to the antipsoriatic agents, betamethasone dipropionate (BD) and salicylic acid (SA), to GsdmA3(Dfl)/+ mice, a hair loss mutant which also exhibits epidermal hyperproliferation akin to psoriasis. The mice were dosed with 100 mg of the test formulation and after 10 days, the mice were sacrificed, skin sections excised and subjected to immunohistochemical determination of COX-2, K17 and MAC-1; and immunofluorescence of Ki-67. Unchanged expression of the proinflammatory enzyme COX-2 was observed in all treatments, suggesting the noninvolvement of COX-2 in the aetiology of cutaneous aberration seen in GsdmA3(Dfl)/+ mice. Intense staining of K17 and MAC-1 in the FO-treated group mirrored the epidermal thickening seen observed in live mice by optical coherence tomography (OCT). The ratio of Ki-67-positive nuclei per 100 basal cells indicated that hyperproliferation of keratinocytes occurred in FO-treated mice and the opposite was true for BD-treated mice. There was a positive correlation (R (2) 0.995) between Ki-67 and the epidermal thickness data observed previously. In all immunochemical procedures, the combined BD, SA and FO formulation did not show any significant difference with the control group, reflecting observations seen previously. In conclusion, the epidermal changes observed following topical FO treatment on GsdmA3(Dfl)/+ mice involves an increase in cellular proliferation and macrophages, although COX-2 does not appear to play an important role.

Matched MeSH terms: Fluorescent Antibody Technique
Nipah Virus Infection of Immature Dendritic Cells Increases Its Transendothelial Migration Across Human Brain Microvascular Endothelial Cells

Tiong V, Shu MH, Wong WF, AbuBakar S, Chang LY

Front Microbiol, 2018;9:2747.
PMID: 30483242 DOI: 10.3389/fmicb.2018.02747

Nipah virus (NiV) can infect multiple organs in humans with the central nervous system (CNS) being the most severely affected. Currently, it is not fully understood how NiV spreads throughout the body. NiV has been shown to infect certain leukocyte populations and we hypothesized that these infected cells could cross the blood-brain barrier (BBB), facilitating NiV entry into the CNS. Here, three leukocyte types, primary immature dendritic cells (iDC), primary monocytes (pMO), and monocytic cell line (THP-1), were evaluated for permissiveness to NiV. We found only iDC and THP-1 were permissive to NiV. Transendothelial migration of mock-infected and NiV-infected leukocytes was then evaluated using an in vitro BBB model established with human brain microvascular endothelial cells (HBMEC). There was approximately a threefold increase in migration of NiV-infected iDC across endothelial monolayer when compared to mock-infected iDC. In contrast, migration rates for pMO and THP-1 did not change upon NiV infection. Across TNF-α-treated endothelial monolayer, there was significant increase of almost twofold in migration of NiV-infected iDC and THP-1 over mock-infected cells. Immunofluorescence analysis showed the migrated NiV-infected leukocytes retained their ability to infect other cells. This study demonstrates for the first time that active NiV infection of iDC and THP-1 increased their transendothelial migration activity across HBMEC and activation of HBMEC by TNF-α further promoted migration. The findings suggest that NiV infection of leukocytes to disseminate the virus via the "Trojan horse" mechanism is a viable route of entry into the CNS.

Matched MeSH terms: Fluorescent Antibody Technique
Fulltext Cytotoxic and apoptosis-inducing effects of wildtype and mutated Hydra actinoporin-like toxin 1 (HALT-1) on various cancer cell lines

Ng TJ, Teo MYM, Liew DS, Effiong PE, Hwang JS, Lim CSY, et al.

PeerJ, 2019;7:e6639.
PMID: 31106043 DOI: 10.7717/peerj.6639

Background: Hydra actinoporin like toxin -1 (HALT-1), is a small 18.5 kDa pore forming toxin derived from Hydra magnipapillata which has been shown to elicit strong haemolytic and cytolytic activity when in contact with cell membranes. Due to its cytotoxic potency, HALT-1 was further investigated for its potential as a toxin moiety candidate in immunotoxin developmental efforts, ideally as a form of targeted therapy against cancer.
Methods: In this study, wtHALT-1 (wild type) and its Y110A mutated binding domain counterpart (mHALT-1) were produced and evaluated for their cytotoxic and apoptotic effects on various cancer cell lines. A total of seven different tumour and non-tumour cell lines including HeLa, HepG2, SW-620, MCF-7, CCD841CoN, NHDF and HCT116 were used. Immunofluorescence assays were used to observe membrane binding and localization changes between both HALT-1 recombinant proteins based on 6xHis-tag detection.
Result: Based on MTT data, mHALT-1 demonstrated a significant reduction of 82% ± 12.21% in cytotoxic activity across all cell lines after the membrane recognition domain had been mutated in comparison to the wtHALT-1. Annexin V FITC/PI assay data also indicated that HeLa, HepG2 and MCF-7 demonstrated an apoptosis-mediated cell death after being treated with wtHALT-1. Additionally, a notable difference between wtHALT-1 and mHALT-1 binding affinity was clearly observed where emission of green fluorescence along the cell membrane was observed only in wtHALT-1 treated cells.
Discussion: These results suggest that mHALT-1 (Y110A) can be potentially developed as a toxin-moiety candidate for the development of future immunotoxins against various human cell-based diseases.

Matched MeSH terms: Fluorescent Antibody Technique
Live-cell imaging and ultrastructural analysis reveal remarkable features of cultured porcine gonocytes

Awang-Junaidi AH, Fayaz MA, Kawamura E, Sobchishin L, MacPhee DJ, Honaramooz A

Cell Tissue Res, 2020 Aug;381(2):361-377.
PMID: 32388763 DOI: 10.1007/s00441-020-03218-5

Gonocytes in the neonatal testis have male germline stem cell potential. The objective of the present study was to examine the behavior and ultrastructure of gonocytes in culture. Neonatal porcine testis cells were cultured for 4 weeks and underwent live-cell imaging to explore real-time interactions among cultured cells. This included imaging every 1 h from day 0 to day 3, every 2 h from day 4 to day 7, and every 1 h for 24 h at days 14, 21, and 28. Samples also underwent scanning electron microscopy, transmission electron microscopy, morphometric evaluations, immunofluorescence, and RT-PCR. Live-cell imaging revealed an active amoeboid-like movement of gonocytes, assisted by the formation of extensive cytoplasmic projections, which, using scanning electron microscopy, were categorized into spike-like filopodia, leaf-like lamellipodia, membrane ruffles, and cytoplasmic blebs. In the first week of culture, gonocytes formed loose attachments on top of a somatic cell monolayer and, in week 2, formed grape-like clusters, which, over time, grew in cell number. Starting at week 3 of culture, some of the gonocyte clusters transformed into large multinucleated embryoid body-like colonies (EBLCs) that expressed both gonocyte- and pluripotent-specific markers. The number and diameter of individual gonocytes, the number and density of organelles within gonocytes, as well as the number and diameter of the EBLCs increased over time (P

Matched MeSH terms: Fluorescent Antibody Technique
Fulltext Hepatitis B Core Antigen in Hepatocytes of Chronic Hepatitis B: Comparison between Indirect Immunofluorescence and Immunoperoxidase Method

Raihan R, Tabassum S, Al-Mahtab M, Nessa A, Jahan M, Shamim Kabir CM, et al.

Euroasian J Hepatogastroenterol, 2015 Jan-Jun;5(1):7-10.
PMID: 29201677 DOI: 10.5005/jp-journals-10018-1120

Background: Hepatitis B virus (HBV) infection has many faces. Precore and core promoter mutants resemble inactive carrier status. The identification of hepatitis B core antigen (HBcAg) in hepatocytes may have variable clinical significance. The present study was undertaken to detect HBcAg in chronic hepatitis B (CHB) patients and to assess the efficacy of detection system by indirect immunofluorescence (IIF) and indirect immunoperoxidase (IIP).

Materials and methods: The study was done in 70 chronic HBV-infected patients. Out of 70 patients, eight (11.4%) were hepatitis B e antigen (HBeAg) positive and 62 (88.57%) were HBeAg negative. Hepatitis B core antigen was detected by indirect immunofluorescence (IIF) and indirect immunoperoxidase (IIP) methods in liver tissue.

Results: All HBeAg positive patients expressed HBcAg by both IIF and IIP methods. Out of 62 patients with HBeAg-negative CHB, HBcAg was detected by IIF in 55 (88.7%) patients and by IIP in 51 (82.26%) patients. A positive relation among viral load and HBcAg detection was also found. This was more evident in the case of HBeAg negative patients and showed a positive relation with HBV DNA levels.

Conclusion: Hepatitis B core antigen can be detected using the IIF from formalin fixed paraffin block preparation and also by IIP method. This seems to reflect the magnitudes of HBV replication in CHB.

How to cite this article: Raihan R, Tabassum S, Al-Mahtab M, Nessa A, Jahan M, Kabir CMS, Kamal M, Aguilar JC. Hepatitis B Core Antigen in Hepatocytes of Chronic Hepatitis B: Comparison between Indirect Immunofluorescence and Immunoperoxidase Method. Euroasian J Hepato-Gastroenterol 2015;5(1):7-10.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
N-cadherin regulates beta-catenin signal and its misexpression perturbs commissural axon projection in the developing chicken spinal cord

Yang C, Li X, Wang C, Fu S, Li H, Guo Z, et al.

J Mol Histol, 2016 Dec;47(6):541-554.
PMID: 27650519

N-cadherin is a calcium-sensitive cell adhesion molecule that plays an important role in the formation of the neural circuit and the development of the nervous system. In the present study, we investigated the function of N-cadherin in cell-cell connection in vitro with HEK293T cells, and in commissural axon projections in the developing chicken spinal cord using in ovo electroporation. Cell-cell connections increased with N-cadherin overexpression in HEK293T cells, while cell contacts disappeared after co-transfection with an N-cadherin-shRNA plasmid. The knockdown of N-cadherin caused the accumulation of β-catenin in the nucleus, supporting the notion that N-cadherin regulates β-catenin signaling in vitro. Furthermore, N-cadherin misexpression perturbed commissural axon projections in the spinal cord. The overexpression of N-cadherin reduced the number of axons that projected alongside the contralateral margin of the floor plate, and formed intermediate longitudinal commissural axons. In contrast, the knockdown of N-cadherin perturbed commissural axon projections significantly, affecting the projections alongside the contralateral margin of the floor plate, but did not affect intermediate longitudinal commissural axons. Taken together, these findings suggest that N-cadherin regulates commissural axon projections in the developing chicken spinal cord.

Matched MeSH terms: Fluorescent Antibody Technique
Diffusion-in-gel enzyme-linked immunosorbent assay, ELISA and IFA test in the detection of malarial antibodies

Kumar GS, Mak JW, Lam PL, Tan MA, Lim PK

Southeast Asian J Trop Med Public Health, 1987 Dec;18(4):502-6.
PMID: 3129797

Malarial antibodies in 80 patients were measured using the diffusion-in-gel enzyme linked immunosorbent assay (DIG-ELISA), enzyme-linked immunosorbent assay (ELISA) and the indirect fluorescent antibody (IFA) test. Good correlations were obtained between all three tests in terms of sensitivity and reliability. DIG-ELISA has the advantage of being a rapid diagnostic tool for the detection of malarial antibodies.

Matched MeSH terms: Fluorescent Antibody Technique
Burkholderia thailandensis whole cell antigen cross-reacts with B. pseudomallei antibodies from patients with melioidosis in an immunofluorescent assay

Puthucheary SD, Anuar AS, Tee TS

Southeast Asian J Trop Med Public Health, 2010 Mar;41(2):395-400.
PMID: 20578523

An immunofluorescent assay (IFAT) using whole cell antigen derived from Burkholderia thailandensis used for detection of total antibodies to Burkholderia pseudomallei, was found to compare favorably with a previous published report on a B. pseudomallei IFAT assay. At a 1:20 cut-off titer, the assay had high sensitivity (98.9%) and satisfactory specificity (92.3%), when tested against sera from 94 patients suspected of melioidosis. Sera from 12 patients with culture proven melioidosis gave absolute concordance with the 2 test antigens. No sera from 50 blood donors had a titer of > or =20. Cross-reactivity with patients' sera positive for Chlamydia, Mycoplasma, Legionella and typhoid was not observed, except for 3 sera from typhus patients and one from a patient with leptospirosis. The major advantage of this assay is that the cultivation and preparation of B. thailandensis as antigen can be carried out in any laboratory with basic microbiological set-up. The serodiagnosis of melioidosis can be made safe for medical laboratory personnel, particularly in B. pseudomallei endemic regions.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Occurrence of Cryptosporidium oocysts in Wrinkled Hornbill and other birds in the Kuala Lumpur National Zoo

Rohela M, Lim YA, Jamaiah I, Khadijah PY, Laang ST, Nazri MH, et al.

Southeast Asian J Trop Med Public Health, 2005;36 Suppl 4:34-40.
PMID: 16438177

The occurrence of a coccidian parasite, Cryptosporidium, among birds in the Kuala Lumpur National Zoo was investigated in this study. A hundred bird fecal samples were taken from various locations of the zoo. Fecal smears prepared using direct smear and formalin ethyl acetate concentration technique were stained with modified Ziehl-Neelsen stain. Samples positive for Cryptosporidium with Ziehl-Neelsen stain were later confirmed using the immunofluorescence technique and viewed under the epifluorescence microscope. Six species of bird feces were confirmed positive with Cryptosporidium oocysts. They included Wrinkled Hornbill (Aceros corrugatus), Great Argus Pheasant (Argusianus argus), Black Swan (Cygnus atratus), Swan Goose (Anser cygnoides), Marabou Stork (Leptoptilos crumeniferus), and Moluccan Cockatoo (Cacatua moluccencis). These birds were located in the aviary and lake, with the Moluccan Cockatoo routinely used as a show bird. Results obtained in this study indicated that animal sanctuaries like zoos and bird parks are important sources of Cryptosporidium infection to humans, especially children and other animals.

Matched MeSH terms: Fluorescent Antibody Technique
Long term maintenance of Toxoplasma gondii (Rh strain) in Vero cell line and use of harvested antigens for immunodiagnosis

Suresh K, Mak JW, Yong HS

Southeast Asian J Trop Med Public Health, 1991 Dec;22 Suppl:124-8.
PMID: 1822869

Thirty in vitro serial passages of Toxoplasman gondii cultures in Vero cell line performed once in every five days had a mean increase in parasite count of 74.4 +/- 14.8 times from that of initial counts. Long term cultures in Vero cell line did not alter the virulence of the parasite. The good correlation (r = 0.99) between the IFA titer and ELISA OD values using the parasite antigens from in vitro sources indicates that long term maintenance of T. gondii in culture does not affect significantly the ability to recognize antibodies to surface and soluble antigens. The results also show that soluble antigens containing host cells can be directly used for immunodiagnostic purposes without purification. The in vitro maintenance of T. gondii is safer and cheaper when compared to the in vivo method.

Matched MeSH terms: Fluorescent Antibody Technique
Serological evidence for a high incidence of transmission of Rickettsia tsutsugamushi in two Orang Asli settlements in Peninsular Malaysia

Brown GW, Robinson DM, Huxsoll DL

Am J Trop Med Hyg, 1978 Jan;27(1 Pt 1):121-3.
PMID: 415625

Two communities of Orang Asli (aborigines) in Peninsular Malaysia were observed for evidence of Rickettsia tsutsugamushi infection over periods of 1-8 mo. Sequential sera were examined for antibody by the indirect immunofluorescence test. The incidence of infection in the two self-selected populations in the two communities was calculated to be 3.9% per month and 3.2% per month.

Matched MeSH terms: Fluorescent Antibody Technique
Comparative evaluation of the indirect immunoperoxidase test for the serodiagnosis of rickettsial disease

Kelly DJ, Wong PW, Gan E, Lewis GE

Am J Trop Med Hyg, 1988 Mar;38(2):400-6.
PMID: 3128129 DOI: 10.4269/ajtmh.1988.38.400

An indirect immunoperoxidase test was compared with an indirect fluorescent antibody test and the Weil-Felix OXK test for serodiagnosis of scrub typhus by measuring the rickettsial antigen specific activity of IgG, IgM, and whole globulin. Acute and convalescent sera from 50 Rickettsia tsutsugamushi isolate-positive scrub typhus patients and from 45 febrile patients diagnosed as having diseases other than scrub typhus were tested. The receiver operating characteristic for each test showed that the indirect immunoperoxidase and indirect fluorescent antibody tests were more sensitive and specific than the Weil-Felix test using convalescent and acute as well as paired sera. The indirect immunoperoxidase test showed no cross-reactivity when R. tsutsugamushi antigen was tested against sera collected from patients living outside the scrub typhus-endemic area with diseases other than scrub typhus. The indirect immunoperoxidase and indirect fluorescent antibody tests were comparable in measured response to R. tsutsugamushi, R. typhi, and TT-118 (spotted fever group) antigen. Thus the indirect immunoperoxidase test represents a sensitive, specific, reproducible, and practical semiquantitative test for rickettsial disease diagnosis.

Matched MeSH terms: Fluorescent Antibody Technique

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