MyMedR

Fulltext Comparison between flowcytometry and immunoperoxidase staining for the enumeration of lymphocyte subsets

Dhaliwal JS, Malar B, Quck CK, Sukumaran KD, Hassan K

Singapore Med J, 1991 Jun;32(3):163-5.

Immunoperoxidase staining was compared with flowcytometry for the enumeration of lymphocyte subsets. The percentages obtained for peripheral blood lymphocytes using immunoperoxidase (CD3 = 76 CD4 = 27.9, B = 10.7 CD4/CD8 = 1.8) differed significantly from those obtained by flowcytometry (CD3 = 65.7 CD4 = 39.4, CD8 = 25.6, B = 16.7, HLA DR = 11.9 CD4/CD8 = 1.54) for certain subsets (CD3, CD4, B). There was no significant difference in lymphocyte subsets between children and adults using the same method. These differences are probably due to the different methods used to prepare lymphocytes for analysis. Other factors that should also be considered are the presence of CD4 antigen on monocytes and CD8 on natural killer cells.

Matched MeSH terms: Immunoenzyme Techniques*

Detection of Malaysian schistosomiasis in Orang Asli of Peninsular Malaysia using serodiagnostic tests

Anuar H, Greer GJ, Ow-Yang CK, Sukumaran KD

Southeast Asian J Trop Med Public Health, 1984 Dec;15(4):479.

PMID: 6398916

Matched MeSH terms: Immunoenzyme Techniques*

Fulltext Human immunodeficiency virus 2 (HIV 2)

How VJ

Malays J Pathol, 1990 Jun;12(1):59-60.

PMID: 2090890

Matched MeSH terms: Immunoenzyme Techniques

Fulltext New sites of localisation of Pasteurella multocida B:2 in buffalo surviving experimental haemorrhagic septicaemia

Annas S, Zamri-Saad M, Jesse FF, Zunita Z

BMC Vet Res, 2014;10:88.

PMID: 24721163 DOI: 10.1186/1746-6148-10-88

Haemorrhagic septicaemia (HS) is an acute septicaemic disease of buffalo and cattle caused by Pasteurella multocida B:2 and E:2. Field outbreaks of HS are known to result in localisation of bacteria in the tonsils of surviving buffalo, confirming that animals can become carriers and the role of respiratory tract in the transmission of the disease. This report describes additional sites of localisation of P. multocida B:2 in surviving buffalo following experimental induction of HS.

Matched MeSH terms: Immunoenzyme Techniques/veterinary

Development of an indirect enzyme immunoassay using monoclonal antibodies for the measurement of 17alpha-hydroxyprogesterone in human serum

Chong H, Cheah SH, Ragavan M, Johgalingam VT

J Immunoassay Immunochem, 2009;30(2):166-79.

PMID: 19330642 DOI: 10.1080/15321810902782863

An indirect enzyme immunoassay for the measurement of total 17alpha-hydroxyprogesterone (17OHP) in serum using monoclonal antibodies generated in our laboratory was developed. Here, (a) instead of extraction with solvents, serum was heated to free protein-bound 17OHP and assay was performed at pH 9.6, (b) to ensure uniform assay conditions for both standards and samples, buffer for standards contained charcoal-stripped pre-heated pooled cord serum. Assays were done in 96-well EIA microplates pre-coated with 17alpha-hydroxyprogesterone-3-(o-carboxymethyl)oxime: bovine serum albumin. Secondary antibody was horseradish peroxidase-linked sheep anti-mouse IgG polyclonal antibody. The method was accurate and suitable for screening for congenital adrenal hyperplasia.

Matched MeSH terms: Immunoenzyme Techniques*

Enzyme immunoassay for the detection of porcine gelatine in edible bird's nests

Tukiran NA, Ismail A, Mustafa S, Hamid M

Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2015;32(7):1023-8.

PMID: 25861981 DOI: 10.1080/19440049.2015.1039605

Porcine gelatine is a common adulterant found in edible bird's nests (EBNs) used to increase the net weight prior to sale. This study aimed to develop indirect enzyme-linked immunosorbent assays (ELISAs) for porcine gelatine adulteration using anti-peptide polyclonal antibodies. Three indirect ELISAs were developed (PAB1, 2 and 3), which had limits of detection (LODs) of 0.12, 0.10 and 0.11 µg g(-1), respectively. When applied to standard solutions of porcine gelatine, the inter- and intra-assays showed coefficients of variation (CVs) less than 20% and were able to detect at least 0.5 ng µg(-1) (0.05%) porcine gelatine in spiked samples. The proposed ELISA offers attractions for quality control in the EBN industry.

Matched MeSH terms: Immunoenzyme Techniques/methods*

Fulltext Dot enzyme immunoassay: an alternative diagnostic aid for dengue fever and dengue haemorrhagic fever

Cardosa MJ, Tio PH

Bull World Health Organ, 1991;69(6):741-5.

PMID: 1786623

A dot enzyme immunoassay (DEIA) for the detection of antibodies to dengue virus was tested for use as a tool in the presumptive diagnosis of dengue fever and dengue haemorrhagic fever. Paired sera from the following groups of patients were tested using the DEIA and the haemagglutination inhibition (HI) test: those with primary dengue fever; those experiencing a second dengue infection; and febrile patients who did not have dengue. The data obtained show that the DEIA can be effectively used at a serum dilution of 1:1000 to confirm presumptive recent dengue in patients with a second dengue infection. However, demonstration of seroconversion proved necessary for patients with primary dengue. At a serum dilution of 1:1000 the DEIA has a specificity of 97.3%. The role of this simple and rapid test in improving the effectivity of programmes for the control of dengue virus infection is discussed.

Matched MeSH terms: Immunoenzyme Techniques*

An in-house assay for serum deoxythymidine kinase

Seah LH, Ton SH, Cheong SK, Hamidah NH

Malays J Pathol, 1991 Dec;13(2):109-13.

PMID: 1823092

An in-house method which utilizes 14C-thymidine as a substrate was used to assay deoxythymidine kinase in serum. The method is sensitive enough to detect normal levels of serum deoxythymidine kinase and the assay procedure also enables rapid handling of multiple samples. With a total reaction volume of 60 ul, the enzyme reaction was found to be linear with concentrations for up to 650 U/L of TK activity. On studying serum deoxythymidine kinase (s-TK) activity with incubation time, there was a proportional increase in activity with the length of incubation time. "Within-batch" precision showed a coefficient of variation (CV) of 4.7% for serum with extremely high s-TK levels and a CV of 8.8% for serum with normal s-TK levels. S-TK showed a CV of less than 16.0% in its activity when stored at -8 degrees C and at -20 degrees C. The normal reference range obtained for s-TK activity was 8.6 +/- 7.5 U/L.

Matched MeSH terms: Immunoenzyme Techniques*

A sensitive malaria immunoperoxidase assay for the detection of Plasmodium falciparum antibody

Lim TS

Am J Trop Med Hyg, 1988 Mar;38(2):255-7.

PMID: 3281491

A new and rapid malaria immunoperoxidase assay using the enzyme horseradish peroxidase in place of fluorescein isothiocyanate was developed to allow the serological measurement of antimalarial antibody by light microscopy. Acetone-fixed thin blood films prepared from cultured Plasmodium falciparum were used as the source of antigen. This malaria immunoperoxidase assay is as sensitive as, and occasionally more sensitive than, the indirect fluorescent antibody assay. It is easy to perform and the antigen used does not show cross-reactivity with sera from nonmalarial diseases.

Matched MeSH terms: Immunoenzyme Techniques*

Comparison of direct immunoperoxidase and direct immunofluorescence for the detection of herpes simplex virus antigen in cell culture

Sabil D, Othman SK, Isahak I

Malays J Pathol, 1990 Jun;12(1):35-8.

PMID: 1965320

150 specimens from suspected herpes simplex genital and skin lesions were received in virus transport medium. They were inoculated into Hep-2 cell monolayers, examined for the presence of virus by cytopathic effect (CPE), direct immunoperoxidase (DIP) and direct immunofluorescence (DIF). Of 39 (26.0%) virus-positive specimens by CPE, 37 (24.7%) were HSV-positive by DIP and 36 (24.0%) by DIF staining. DIP staining had a sensitivity of 100%, specificity of 99.1%, positive predictive value of 97.3% and negative predictive value of 100% in relation to DIF as a standard test. Of 39 specimens positive by CPE, only 25.6% were HSV-positive within 24 h post-inoculation compared to 94% HSV-positive by DIP and DIF staining at the same time.

Matched MeSH terms: Immunoenzyme Techniques*

Development of a dot enzyme immunoassay for dengue 3: a sensitive method for the detection of antidengue antibodies

Cardosa MJ, Hooi TP, Shaari NS

J Virol Methods, 1988 Oct;22(1):81-8.

PMID: 3058737

Partially purified DEN3 virus was used as antigen in a sensitive dot enzyme immunoassay (DEIA) for the detection of antibodies to flavivirus antigens. We describe here the method used to prepare and optimise the antigen-bearing nitrocellulose membranes and present the results obtained from screening 20 acute phase sera from patients shown to have had recent dengue infections by the haemagglutination inhibition (HI) test. Sixteen pairs of acute and convalescent sera from dengue-negative patients had no detectable antibody to dengue virus by HI. These were shown to have no antibody detectable by DEIA. Sera positive for dengue antibodies by HI had DEIA titers ranging from 10 to several thousand times greater than the titers detected by HI.

Matched MeSH terms: Immunoenzyme Techniques*

Detection of rickettsial antibodies using Weil-Felix (OXK and OX19) antigens and the indirect immunoperoxidase assay

Tay ST, Kamalanathan M, Rohani MY

Southeast Asian J Trop Med Public Health, 2003 Mar;34(1):171-4.

PMID: 12971531

Matched MeSH terms: Immunoenzyme Techniques*

Fulltext A measles IgM rapid diagnostic test to address challenges with national measles surveillance and response in Malaysia

Senin A, Noordin NM, Sani JAM, Mahat D, Donadel M, Scobie HM, et al.

PLoS One, 2024;19(3):e0298730.

PMID: 38483868 DOI: 10.1371/journal.pone.0298730

INTRODUCTION: A lateral flow rapid diagnostic test (RDT) enables detection of measles specific immunoglobulin M (IgM) antibody in serum, capillary blood, and oral fluid with accuracy consistent with enzyme immunoassay (EIA). The objectives of the study were: 1) to assess measles RDT inter-reader agreement between two clinic staff; 2) to assess the sensitivity and specificity of the measles RDT relative to standard surveillance testing in a low transmission setting; 3) to evaluate the knowledge, attitudes, and practices of staff in clinics using the RDT; and 4) to assess the impact of RDT testing on the measles public health response in Malaysia.

MATERIALS AND METHODS: The clinic-based prospective evaluation included all suspected measles cases captured by routine measles surveillance at 34 purposely selected clinics in 15 health districts in Malaysia between September 2019 and June 2020, following day-long regional trainings on RDT use. Following informed consent, four specimens were collected from each suspected case, including those routinely collected for standard surveillance [serum for EIA and throat swabs for quantitative reverse transcriptase polymerase chain reaction (RT-qPCR)] together with capillary blood and oral fluid tested with RDTs during the study. RDT impact was evaluated by comparing the rapidity of measles public health response between the pre-RDT implementation (December 2018 to August 2019) and RDT implementation periods (September 2019 to June 2020). To assess knowledge, attitudes, and practices of RDT use, staff involved in the public health management of measles at the selected sites were surveyed.

RESULTS: Among the 436 suspect cases, agreement of direct visual readings of measles RDT devices between two health clinic staff was 99% for capillary blood (k = 0.94) and 97% for oral fluid (k = 0.90) specimens. Of the total, 45 (10%) were positive by measles IgM EIA (n = 44, including five also positive by RT-qPCR) or RT-qPCR only (n = 1), and 38 were positive by RDT (using either capillary blood or oral fluid). Using measles IgM EIA or RT-qPCR as reference, RDT sensitivity using capillary blood was 43% (95% CI: 30%-58%) and specificity was 98% (95% CI: 96%-99%); using oral fluid, sensitivity (26%, 95% CI: 15%-40%) and specificity (97%, 95% CI: 94%-98%) were lower. Nine months after training, RDT knowledge was high among staff involved with the public health management of measles (average quiz score of 80%) and was highest among those who received formal training (88%), followed by those trained during supervisory visits (83%). During the RDT implementation period, the number of days from case confirmation until initiation of public response decreased by about 5 days.

CONCLUSION: The measles IgM RDT shows >95% inter-reader agreement, high retention of RDT knowledge, and a more rapid public health response. However, despite ≥95% RDT specificity using capillary blood or oral fluid, RDT sensitivity was <45%. Higher-powered studies using highly specific IgM assays and systematic RT-qPCR for case confirmation are needed to establish the role of RDT in measles elimination settings.

Matched MeSH terms: Immunoenzyme Techniques

Fulltext A case study of human immunodeficiency virus with positive seroconversion to negative

Paranthaman V, Yip HL, Ker HB

Malays Fam Physician, 2015;10(1):44-6.

PMID: 26425294 MyJurnal

This case study demonstrates a 36-year-old ex-intravenous drug user (IVDU) who had been initially tested positive for human immunodeficiency virus (HIV) twice using Enzyme Immunoassay (EIA) method (Particle agglutination, PA done), but a year later he was tested HIV-negative. The patient was asymptomatic for HIV and T helper cells (CD4) count remained stable throughout this period. In light of this case, there may be a need to retest by molecular methods for high risk category patients who were initially diagnosed HIV-positive, but later showing an unexpected clinical course, such as a rising or stable CD4 titre over the years.

Matched MeSH terms: Immunoenzyme Techniques

Rapid thin-layer chromatographic semiquantification of morphine in urine via dabsylation: comparison with EMIT and thin-layer chromatographic-iodoplatinate methods

Wang SY, Tham SY, Poon MK

Clin Chem, 1989 Feb;35(2):329-30.

PMID: 2644060

Matched MeSH terms: Immunoenzyme Techniques

Multicentre evaluation of dengue IgM dot enzyme immunoassay

Lam SK, Fong MY, Chungue E, Doraisingham S, Igarashi A, Khin MA, et al.

Clin Diagn Virol, 1996 Nov;7(2):93-8.

PMID: 9137865 DOI: 10.1016/S0928-0197(96)00257-7

The traditional methods used in the diagnosis of dengue infection do not lend themselves to field application. As such, clinical specimens have to be sent to a central laboratory for processing which invariably leads to delay. This affects patient management and disease control. The development of the dengue IgM dot enzyme immunoassay has opened up the possibility of carrying out the test in peripheral health settings.

Matched MeSH terms: Immunoenzyme Techniques*

Use of avidin-biotin-glucose oxidase complex to detect antimalarial antibody in serum by light microscopy

Lee M, Lambros C

Am J Trop Med Hyg, 1988 Aug;39(2):145-9.

PMID: 3044152

An immunohistochemical assay was developed combining an avidin-biotin-glucose oxidase complex procedure (ABC-GO) with light microscopy to detect specific antibody against Plasmodium falciparum. Thin blood films were prepared from culture material of P. falciparum and fixed with acetone. Antibody was detected by successive incubations with test serum, biotinylated goat antihuman antibody, avidin-biotin-glucose oxidase complex, and glucose oxidase substrate. In the presence of reactive serum, a blue precipitate formed on the parasites and could be visually observed with a 40x objective. Sera from patients with single infections for P. vivax or P. ovale were unreactive. No cross-reactivity was observed with sera from patients with rheumatoid arthritis, filariasis, amebiasis, schistosomiasis, dengue, scrub typhus, leptospirosis, or toxoplasmosis. The sensitivity of ABC-GO is comparable to that of the indirect fluorescent antibody test.

Matched MeSH terms: Immunoenzyme Techniques*

An immunohistochemical method for the diagnosis of melioidosis

Wong KT, Vadivelu J, Puthucheary SD, Tan KL

Pathology, 1996 May;28(2):188-91.

PMID: 8743829

In order to assess the usefulness of immunohistochemistry in the diagnosis of melioidosis, an infection by Burkholderia pseudomallei, polyclonal antibodies were applied to tissues from known cases of melioidosis and to other infected tissues. Formalin-fixed, paraffin-embedded tissues were stained by a modified immunoperoxidase technique. In autopsy tissues with inflammatory lesions of melioidosis, the cytoplasm of phagocytes and intact bacilli, both intra- and extracellular, were stained very strongly positive. Relatively more focal positive staining was observed in some but not all surgical biopsies from proven cases of melioidosis. In granulomas staining was mainly found in the central necrotic areas, with little staining of peripheral phagocytes. All control materials stained negative. Immunohistochemistry appears to be a useful diagnostic tool in melioidosis.

Matched MeSH terms: Immunoenzyme Techniques*

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: Immunoenzyme Techniques*

Infectivity titration of the fast-replicating and cytopathic hepatitis A virus strain HM175A.2 by an in situ enzyme immunoassay

Yap KL, Lam SK

J Virol Methods, 1994 Apr;47(1-2):217-26.

PMID: 8051228

A simple, rapid and objective infectivity assay based on an in situ enzyme immunoassay (EIA) was developed for the fast-growing and cytopathic cell culture-adapted hepatitis A virus (HAV) strain HM175A.2. Infectivity titration by EIA correlated well with titration by cytopathic effects. The reliability of this assay was demonstrated by close agreement in virus infectivity titers among different assays of the same virus aliquot and between assays of different virus aliquots. HAV infected cell cultures after fixation could be stored for up to 1 week before testing without decline in virus titer.

Matched MeSH terms: Immunoenzyme Techniques*

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