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THE INDIRECT FLUORESCENT ANTIBODY TECHNIQUE IN SCRUB TYPHUS STUDIES

ELLISON DW, BAKER HJ

Med J Malaysia, 1964 Sep;19:65-6.
PMID: 14244224
Matched MeSH terms: Fluorescent Antibody Technique, Indirect*
Antinuclear antibody titration and pattern are helpful in the diagnosis of systemic autoimmune rheumatic diseases

Mohd Razi MS, Sugumaran Y, Mohd Haniz NA, Khilmie K, Osmera AH, Jauhary EJ, et al.

Malays J Pathol, 2024 Apr;46(1):63-69.
PMID: 38682845

INTRODUCTION: Anti-nuclear antibody (ANA) testing is among the most common immunological test requested in the diagnostic immunology laboratory. The main purpose of this test is to screen for the underlying systemic autoimmune rheumatic diseases (SARDs). The gold standard laboratory method for ANA detection is by the indirect immunofluorescence (IIF) assay. In most laboratories, positive ANA-IIF is reported in terms of titration and pattern.
OBJECTIVE: This study was conducted with the aim of determining the correlation between ANA-IIF titration and pattern for the diagnosis of SARDs.
MATERIALS AND METHODS: A retrospective study was conducted whereby the positive ANA-IIF samples from 1st July 2018 until 31st December 2019 and 1st January 2021 until 31st March 2021 were included in this study. The duplicate samples were excluded. ANA-IIF titration and pattern were recorded for all patients. The demographic, clinical, and final diagnosis data were retrieved from each patient's clinical note.
RESULTS: A total of 179 patients were included for analysis. The majority of the patients were female (79.9%) and from Malay ethnicity (66.5%). Sixty-five patients (36.3%) had ANA-IIF positive at 1:80 titration followed by 45 patients (25.1%) positive at titration of equal or more than 1:160. Speckled was the predominant pattern visualised in 90 patients (50.3%) followed by homogeneous in 76 patients (42.5%). Forty-five patients (25.1%) were finally diagnosed with SARDs with 41 of them diagnosed as SLE. ANA titration was significantly associated with the final diagnosis of SARDs at all titres (p<0.001) but the best cut-off was noted at a titre of equal or more than 1:320 with the sensitivity and specificity of 86.7% and 77.6% respectively. The homogeneous pattern was also significantly associated with SARDs (p=0.04). The final diagnosis of SARDs were significantly higher in female (p=0.03) and their age was significantly younger (p<0.001).
CONCLUSION: ANA-IIF titration of equal or more than 1:320 can be used as the best titration for differentiating between SARDs and non-SARDs in a positive ANA sample. Patients with homogeneous pattern were more likely to be diagnosed with SARDs than other ANA-IIF patterns.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect/methods
Fulltext The seroprevalence of bovine babesiosis in Malaysia

Rahman WA, Lye YP, Chandrawathani P

Trop Biomed, 2010 Aug;27(2):301-7.
PMID: 20962729 MyJurnal

One hundred sera of Malaysian cattle were used in this seroprevalence study for bovine babesiosis. All sera were obtained from the Serological Unit of the Veterinary Research Institute (VRI), Ipoh, Perak. The sera were tested using a Veterinary Medical Research & Development (VMRD) commercial Indirect Immunofluourescent Antibody Test (IFAT) kit. The results showed that 17.0% were found to be positive for Babesia bovis, 16.0% for Babesia bigemina, and 9.0% for both B. bovis and B. bigemina infections.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Fulltext Indirect immunofluorescence screening of potential arthritic autoantibodies in systemic lupus erythematosus

Selvaraja M, Che Ku Daud CKD, Abdul Jalil M, Md. Shah A, Amin Nordin S, Ahmad Bajari Z, et al.

Malaysian Journal of Medicine and Health Sciences, 2018;14(103):1-4.
MyJurnal

Joint involvement is common in systemic lupus erythematosus (SLE) patients, however, screening for joint specific autoantibodies in patients is not routinely performed. This may be due to the lack of known antigens and available tissue. The rat musculoskeletal tissue may be a suitable source of antigen to detect arthritic autoantibodies.
Method: We tested plasma of SLE patients, with arthritis (N=9) and without arthritis (N=7) as well as plasma from normal individuals (N=7) on fresh sectioned tissue from rat plantar hind paw using indirect immunofluorescence method.
Results: Binding of autoantibodies to striation in skeletal muscle cells in the tissue was clearly demonstrable in all samples from SLE with arthritis but not on slides incubated with plasma from normal or SLE without arthritis.
Conclusion: Thus, rat plantar tissue may be suitable for detecting autoantibodies from SLE patients that may be involved in the pathogenesis of lupus arthritis.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Fulltext Effect of protein concentration and injection pressure in microinjection delivery of maltose binding protein into breast cancer cells

Lim, S.N., Zeenathul, N.A., Mohd Azmi, M.L., Abas Mazni, O., Fauziah, O.

Pertanika Journal of Science & Technology, 2011;19(2):273-283.
MyJurnal

Microinjection is a powerful tool to deliver various substances, such as nucleic acids, proteins, peptides, RNA, and synthetic molecules into mammalian cells mechanically. Through microinjection, a controlled amount of protein can be delivered into the target cells to elucidate the specific functional
effects in vitro. In this study, a series of protein microinjection optimization was performed in human breast cancer cells. The presence of Maltose Binding Protein (MBP) was microscopically monitored through indirect immunofluorescence assay. The optimization experimentation gave a high success rate when MBP protein was used at the minimum concentration of 1.5 mg/ml and at the injection pressures of 50 and 70 hPa. The average success rate of injections was 49.2±4.15% and 50.8±4.6%, while the average cell survivability was 50.98±4.67% and 49.72±5.48% at 50 and 70 hPa, respectively. The optimization of the MBP concentration and injection pressures successfully allowed an efficient delivery of precise protein dosage into breast cancer cells without any adverse effect. This microinjection optimization can be a practical guideline in any downstream applications of protein functional work.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Fulltext Lignosus rhinocerus (Cooke) Ryvarden: A Medicinal Mushroom That Stimulates Neurite Outgrowth in PC-12 Cells

Eik LF, Naidu M, David P, Wong KH, Tan YS, Sabaratnam V

Evid Based Complement Alternat Med, 2012;2012:320308.
PMID: 22203867 DOI: 10.1155/2012/320308

A national treasure mushroom, Lignosus rhinocerus, has been used to treat variety of ailments by local and indigenous communities in Malaysia. The aim of this study was to investigate the potential of the most valuable part of L. rhinocerus, the sclerotium, on neurite outgrowth activity by using PC-12Adh cell line. Differentiated cells with one thin extension at least double the length of the cell diameter were scored positive. Our results showed that aqueous sclerotium L. rhinocerus extract induced neurite outgrowths of 24.4% and 42.1% at 20 μg/mL (w/v) of aqueous extract alone and a combination of 20 μg/mL (w/v) aqueous extract and 30 ng/mL (w/v) of NGF, respectively. Combination of NGF and sclerotium extract had additive effects and enhanced neurite outgrowth. Neuronal differentiation was demonstrated by indirect immunofluorescence of neurofilament protein. Aqueous sclerotium extract contained neuroactive compounds that stimulated neurite outgrowth in vitro. To our knowledge this is the first report on neurite-stimulating activities of L. rhinocerus.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Diagnostic and prognostic value of an immunofluorescent assay for melioidosis

Vadivelu J, Puthucheary SD

Am J Trop Med Hyg, 2000 Feb;62(2):297-300.
PMID: 10813488

Melioidosis caused by Burkholderia pseudomallei is endemic in southeast Asia. The clinical manifestations range from wound infections to acute septicemia. In some cases, recurrence can also occur following complete recovery. Case fatality rates are high and a major factor is the delay in the culture and identification of the bacterium. An immunofluorescent assay (IFAT) using whole-cell antigen for the detection of total antibodies to B. pseudomallei was tested with 650 sera. Using a cut-off value of 1:80, 66 sera from culture-confirmed cases were positive with titers > or = 320. In another 523 sera from patients in which no other etiology could be found, 149 (23.4%) were positive. To monitor disease activity, persistence of antibody levels was investigated on 61 serial sera samples collected from 14 other confirmed cases on follow-up visits while on oral maintenance therapy. The IFAT demonstrated a reduction in titers in cases of localized infections, suggesting that either the infection was being resolved or arrested while septicemic patients maintained high IFAT titers on follow-up, suggesting the possibility of continuous sequestration of antigen from an intracellular source.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect*
Fulltext Serodiagnosis of melioidosis in Malaysia

Vadivelu J, Puthucheary SD, Gendeh GS, Parasakthi N

Singapore Med J, 1995 Jun;36(3):299-302.
PMID: 8553097

Current diagnosis of melioidosis is based on bacterial culture and/or serology which is becoming increasingly useful. An IgM-ELISA using heat-killed whole cells of Pseudomonas pseudomallei was developed and compared with an indirect haemagglutination technique (IHAT) and an indirect immunofluorescent technique(IFAT). The IgM-ELISA using a P:N ratio of > or = 2 had a sensitivity of 91% and a specificity of 96%. All 3 assays were further used in a seroepidemiological survey amongst different groups of patients and healthy individuals. It was found that the IFAT performed better than the IHAT, detecting antibodies to P. pseudomallei in 6% of diabetics, 5% of pyrexics, 8% of pregnant women and 3% of farmers. For the same groups the IgM-ELISA detected antibodies in 1% of pyrexics, 8% of pregnant women and a further 14% of farmers. The IgM-ELISA was found to be sensitive and useful for the serological diagnosis of acute melioidosis.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect/methods*
Establishment of an immunofluorescence assay to detect IgM antibodies to Nipah virus using HeLa cells expressing recombinant nucleoprotein

Kaku Y, Park ES, Noguchi A, Inoue S, Lunt R, Malbas FF, et al.

J Virol Methods, 2019 07;269:83-87.
PMID: 30954461 DOI: 10.1016/j.jviromet.2019.03.009

A novel indirect fluorescent antibody test (IFAT) for detection of IgM against Nipah virus (NiV) was developed using HeLa 229 cells expressing recombinant NiV nucleocapsid protein (NiV-N). The NiV IFAT was evaluated using three panels of sera: a) experimentally produced sera from NiV-N-immunized/pre-immunized macaques, b) post-infection human sera associated with a Nipah disease outbreak in the Philippines in 2014, and c) human sera from a non-exposed Malaysian population. Immunized macaque sera showed a characteristic granular staining pattern of the NiV-N expressed antigen in HeLa 229 cells, which was readily distinguished from negative-binding results of the pre-immunized macaque sera. The IgM antibody titers in sequential serum samples (n = 7) obtained from three Nipah patients correlated well with previously published results using conventional IgM capture ELISA and SNT serology. The 90 human serum samples from unexposed persons were unreactive by IFAT. The IFAT utilizing NiV-N-expressing HeLa 229 cells to detect IgM antibody in an early stage of NiV infection is an effective approach, which could be utilized readily in local laboratories to complement other capabilities in NiV-affected countries.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect/methods*
First report of Cryptosporidium deer-like genotype in Malaysian cattle

Halim NA, Plutzer J, Bakheit MA, Karanis P

Vet Parasitol, 2008 Apr 15;152(3-4):325-9.
PMID: 18289793 DOI: 10.1016/j.vetpar.2007.12.035

Fifty faecal samples from diarrheic calves between 1 and 6 months old were collected per rectum from 5 farms around Petaling District in Selangor, Malaysia for Cryptosporidium species detection and genotyping investigation. Oocysts were purified using sedimentation and gradient centrifugation, then examined by immunofluorescence assay (IFAT). Genomic DNA was extracted from all samples and nested PCR was performed to amplify the SSU rRNA gene. Eighteen samples (36%) were positive for Cryptosporidium species by PCR. The sequence and phylogenetic analysis of 14 isolates indicated that Cryptosporidium parvum was most common (11 isolates) followed by Cryptosporidium deer-like genotype (3 isolates). The present work reports the first data on Cryptosporidium genotyping from cattle in Malaysia.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect/veterinary
Detection of IgM antibodies against Legionella pneumophila serogroup 1 in Malaysian blood donors and patients with respiratory illnesses: evaluation of enzyme-linked immunosorbent assay and indirect immunofluorescence assay

Tay ST, Lakhbeer Singh HK, Ramasame SD, Vadivelu J

Jpn J Infect Dis, 2009 Sep;62(5):409-10.
PMID: 19762997
Matched MeSH terms: Fluorescent Antibody Technique, Indirect/methods*
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
Fulltext Antibodies in systemic lupus antineutrophil cytoplasmic erythematosus: prevalence, disease activity correlations and organ system associations

Fauzi AR, Kong NCT, Chua MK, Jeyabalan V, Idris MN, Azizah R

Med J Malaysia, 2004 Aug;59(3):372-7.
PMID: 15727383

Systemic Lupus Erythematosus (SLE) is a disease with multiorgan involvement and multiple autoantibody production including antineutrophil cytoplasmic antibodies (ANCA). Despite its reported prevalence in more than one third of SLE patients, the role of ANCA in the pathogenesis or otherwise in SLE remains unresolved. 131 SLE patients had been previously studied for various serologic parameters of disease activity. Their cumulative organ involvement in the course of their disease had also been determined and the Lupus Activity Index (LAI) calculated. Their stored sera were then screened for the presence of ANCA by two methods viz Indirect immunofluorescence (IIF) and also enzyme-linked immunosorbent assay (ELISA). ANCA was present in 24.8% of these SLE patients. The atypical ANCA pattern was predominant and accounted for an overall of 20.6%. Anti-MPO and anti-PR3 were detected in 1.5% of patients respectively. No association was found between ANCA positivity and disease activity. There was also no association of ANCA with specific organ involvement. Despite the high prevalence of ANCA especially the atypical variant in SLE, they probably represent only one of the wide repertoire of autoantibodies found in this disease. Routine testing for ANCA in lupus patients is therefore not recommended.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Anti-tumour promoter activity in Malaysian ginger rhizobia used in traditional medicine

Vimala S, Norhanom AW, Yadav M

Br. J. Cancer, 1999 Apr;80(1-2):110-6.
PMID: 10389986

Zingiberaceae rhizomes commonly used in the Malaysian traditional medicine were screened for anti-tumour promoter activity using the short-term assay of inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced Epstein-Barr virus early antigen (EBV-EA) in Raji cells. The inhibition of TPA-induced EBV-EA was detected using the indirect immunofluorescence assay (IFA) and Western blot technique. The indirect IFA detected the expression/inhibition of EBV-EA-D (diffused EA antigen), whereas the Western blot technique detected the expression/inhibition of both EBV-EA-D and EA-R (restricted EA antigen). Seven rhizomes were found to possess inhibitory activity towards EBV activation, induced by TPA; they are: Curcuma domestica, C. xanthorrhiza, Kaempferia galanga, Zingiber cassumunar, Z. officinale, Z. officinale (red variety), and Z. zerumbet. A cytotoxicity assay was carried out to determine the toxicity of the Zingiberaceae rhizome extracts. The rhizome extracts that exhibited EBV activation inhibitory activity had no cytotoxicity effect in Raji cells. Therefore, the present study shows that several Zingiberaceae species used in Malaysian traditional medicine contain naturally occurring non-toxic compounds that inhibit the EBV activation, which, if further investigated, could contribute in the development of cancer prevention methods at the tumour-promoting stage.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Fulltext Detection of anti-cell membrane DNA antibodies by indirect immunoflouresencence technique in systemic lupus erythematosus

Faten Nurul Amira Awing Kechik, Maha Abdullah, Masriana Hassan, Masita Arip, Hasni Mahayidin

Malaysian Journal of Medicine and Health Sciences, 2019;15(108):28-28.
MyJurnal

Introduction: Systemic lupus erythematosus (SLE) has a broad spectrum of clinical presentations. The diagnosis of SLE remains a challenge and largely depends on the presence of several serum autoantibodies including anti-nuclear antibody (ANA), anti-double-stranded DNA antibody (anti-dsDNA) and anti-Smith antibody (anti-Sm). ANA, a highly sensitive but not specific marker is used for SLE screening Anti-dsDNA and anti-Sm are SLE-specific biomarkers but has lower sensitivity of 80% and 30% for SLE, respectively. However, it is noted that there are SLE patients who are persistently negative for SLE-specific autoantibodies. Anti-dsDNA and anti-Sm were reported to be negative in up to 51.2% and 62.4% of SLE, respectively. This limitation can lead to misdiagnosis and halter proper treatment to SLE patients. Previous studies have suggested that cell membrane DNA (cmDNA) can act as a specific target for the autoantibodies in SLE patients. Autoantibodies towards cmDNA (anti-cmDNA) were reported to have promis-ing value as a reliable biomarker for SLE. In this study, we would like to determine the usefulness of anti-cmDNA in diagnosing SLE as compared to the standard SLE-specific autoantibodies. Methods: Serum samples from 83 SLE patients, 86 other connective tissue diseases and 61 healthy subjects were included in this study. The other connec-tive tissue diseases include samples from 10 Sjogren’s syndrome, 56 rheumatoid arthritis, 12 scleroderma and eight mixed connected tissues disease (MCTD) patients. All samples were analysed by indirect immunofluorescence (IIF) technique using Raji cells as substrate to detect the presence of anti-cmDNA. Anti-cmDNA was reported as positive if there was presence of a fluorescent ring, either continuous or punctate. Sera from SLE patients were also tested for anti-dsDNA and anti-Sm antibodies by using enzyme-immunoassays. Results: Anti-cmDNA positivity was highest in SLE (55.4%) than in other connective tissue diseases (9.3%) and healthy subjects (0%). Anti-cmDNA was 100% spe-cific at differentiating SLE from healthy subjects and 90.7% specific at differentiating SLE from other connective tissue diseases. There was no difference in the sensitivity (55.4%) of anti-cmDNA at differentiating SLE from both groups. Anti-cmDNA were present in 46 SLE samples negative for standard SLE-specific autoantibodies. It was detected in 11 (42.3%) of anti-dsDNA, 23 (63.9%) of anti-Sm and 8 (12.9%) of both anti-Sm and anti-dsDNA negative samples. Conclusion: The high specificity of anti-cmDNA detection using IIF method makes it an excellent diagnostic tool for SLE. Anti-cmDNA is potentially a very useful biomarker for SLE with negative anti-dsDNA or/and anti-Sm antibodies.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
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
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
Isolation of Neospora caninum from a calf in Malaysia

Cheah TS, Mattsson JG, Zaini M, Sani RA, Jakubek EB, Uggla A, et al.

Vet Parasitol, 2004 Dec 15;126(3):263-9.
PMID: 15567590

In order to attempt isolate the protozoan parasite Neospora caninum, an N. caninum seropositive pregnant Sahiwal Friesian cross heifer from a large-scale dairy farm in Malaysia was kept for observation until parturition at the Veterinary Research Institute, Ipoh. The heifer gave birth to a female calf that was weak, underweight and unable to rise. Precolostral serum from the calf had an N. caninum indirect fluorescent antibody test titre of 1:3200. It died 12 h after birth and necropsy was performed. Brain homogenate from the calf was inoculated into 10 BALB/c mice that were kept for 3 months after which brain tissue from the mice was inoculated onto 24 h fresh monolayer Vero cell lines. The cell cultures were examined daily until growth of intracellular protozoa was observed. DNA of the organisms from the cell cultures was analyzed by PCR and DNA sequencing. DNA fragments of the expected size were amplified from the isolate using N. caninum-specific primers, and sequence analysis of ITS1 clearly identified the isolate as N. caninum. This is the first successful isolation of N. caninum from a bovine in Malaysia, and the isolate is designated Nc-MalB1.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect/veterinary
Fulltext Expression of survivin in fetal and adult normal tissues of rat

Iskandar ZA, Al-Joudi FS

Malays J Pathol, 2006 Dec;28(2):101-5.
PMID: 18376799 MyJurnal

Survivin is an inhibitor of apoptosis protein and regulates the cell cycle in the G2/M phase. Survivin is expressed during embryonic and fetal development, selectively over-expressed in common human cancers and completely down-regulated in normal adult tissue. This work was aimed at studying the expression of the survivin homologues and their subcellular distribution in fetal and normal adult tissues of rat. Survivin expression was evaluated by immunohistochemistry in formalin-fixed, paraffin-embedded tissue sections of fetal and normal adult tissues of rat using the polyclonal serum SUR12A-CFI. This serum demonstrated intense positive survivin staining in adult kidney, ovary and oviduct, and a variable expression in different fetal organs, with particularly intense expression detected in the adrenal gland, liver, stomach, small intestine, colon, kidney and skin. In both fetal and adult tissues, the expression was predominantly cytoplasmic. It was concluded that survivin was abundantly and prominently expressed during fetal development in rat and that the polyclonal anti-human survivin antibody SUR12A-CFI is reactive with rat survivin.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect
Comparison of two monoclonal antibody kits with cell culture isolation in the detection of respiratory virus antigens

Khairullah NS

Malays J Pathol, 1996 Jun;18(1):27-30.
PMID: 10879221

Two different preparations of monoclonal antibodies developed against respiratory viruses have been evaluated by the immunofluorescence antibody technique. The Chemicon monoclonal antibodies were found to be more efficient at picking up positive specimens with a high sensitivity and specificity than Imagen monoclonal antibodies. However, the overall concordance rate of the monoclonal antibodies was 92.3%-100%. Generally, when compared with cell culture isolation, the immunofluorescence antibody technique was found to be more sensitive. The high quality of the Chemicon monoclonal antibodies contribute to their value in providing definitive diagnosis, within a few hours of specimen collection, thus allowing early management of patients, their contacts and control of hospital infection.

Matched MeSH terms: Fluorescent Antibody Technique, Indirect

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