Recombinant leptospiral immunoglobulin like B protein based latex agglutination test for serodiagnosis of human leptospirosis

Humans get leptospirosis by contact with fresh water, damp soil, or vegetation contaminated by the urine of infected animals, swallowing contaminated food or water or while working in contaminated flood plains or at wet agricultural settings. The bacteria enter the body through abrasions in the skin and mucous membranes. In the present study recombinant LigB protein is employed in latex agglutination test, which is a cross reacting lipoprotein able to detect acute infection caused by any pathogenic leptospiral serovars. It was employed for serodiagnosis of leptospirosis. The 46 KDa 6X His tagged LigB protein, obtained by IPTG induction of recombinant E. coli M15 cells containing the N-terminal region of LigB gee in PQE30 expression vector, was purified by Ni-NTA affinity chromatography and adsorbed on latex bead surface for performing latex agglutination test against leptospirosis suspected human sera. A total of 28 human sera samples were received from Post Graduate Institute of Medical Education & Research (PGIMER), Chandigarh India, which tested positive by IgM ELISA test kit were subjected to both rLigB based LAT and MAT. All the 28 sera showed seropositivity by both the tests. Icterohaemorrahigae was the predominant serovar followed by Javanica and Grippotyphosa. Six out seven sera samples received from Indian Veterinary research Institute, Human Hospital and City Hospital, Bareilly were tested positive both by rLAT and MAT. The result showed that sera were tested positive by rLigB based LAT, which were reconfirmed using microscopic agglutination test (MAT). The results from LAT were in concordance with MAT. In conclusion, rLigB based LAT is a rapid, reliable diagnostic tool at resource poor and remote diagnostic centers with high sensitivity and specificity, under laboratory and field conditions, for the detection of leptospirosis. Asian J. Med. Biol. Res. June 2020, 6(2): 229-236

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Cloning and Expression of Leptospiral Immunoglobulin Like B Gene and Use of The Recombinant Antigen for The Diagnosis of Bovine and Caprine Leptospirosis

International Journal of Applied Sciences and Biotechnology ◽

10.3126/ijasbt.v8i2.29588 ◽

2020 ◽

Vol 8 (2) ◽

pp. 146-153

Author(s):

Yosef Deneke ◽

Rajib Deb

Keyword(s):

Acute Infection ◽

High Sensitivity ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Cloning And Expression ◽

Indian Veterinary Research Institute ◽

Iptg Induction ◽

E Coli ◽

Veterinary Research

In the present study recombinant LigB protein is employed in latex agglutination test, which is a cross reacting lipoprotein able to detect acute infection caused by any pathogenic leptospiral serovars. It was employed for serodiagnosis of leptospirosis. The 46KDa 6X His tagged LigB protein, obtained by IPTG induction of recombinant E. coli M15 cells containing the N-terminal region of LigB gee in PQE30 expression vector, was purified by Ni-NTA affinity chromatography and adsorbed on latex bead surface for performing latex agglutination test against Leptospirosis suspected field sea. Western blot confirmed that rLigB is an immunodominant protein against which antibodies are produced during active infection. A total of 453 field sera, including 432 bovine sera, 18 caprine sera and three sera samples of buffalo bull collected post-mortem following death the animal from Indian Veterinary Research institute (IVRI) were tested using rLigB based LAT. The result showed that 300 sera were tested positive by rLigB based LAT, which were reconfirmed using microscopic agglutination test (MAT). The results from LAT were in concordance with MAT. In conclusion, under laboratory and field conditions, rLigB based LAT is a rapid, pen site, reliable diagnostic tool of high sensitivity and specificity for the detection of Leptospirosis. Int. J. Appl. Sci. Biotechnol. Vol 8(2): 146-153

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Comparative evaluation of recombinant LigB based latex agglutination test with microscopic agglutination test for the diagnosis of wildlife leptospirosis

Asian Journal of Medical and Biological Research ◽

10.3329/ajmbr.v6i3.49792 ◽

2020 ◽

Vol 6 (3) ◽

pp. 440-448

Author(s):

Yosef Deneke ◽

Rajib Deb ◽

SM Lutful Kabir

Keyword(s):

Comparative Evaluation ◽

Acute Infection ◽

High Sensitivity ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Microscopic Agglutination Test ◽

Iptg Induction ◽

E Coli ◽

Sera Samples

In the present study a comparative evaluation microscopic agglutination test with rLigB protein based latex agglutination test was carried out, which is a cross reacting lipoprotein able to detect acute infection caused by any pathogenic leptospiralserovars. It was employed for serodiagnosis of leptospirosis. The 46 KDa 6X His tagged LigB protein, obtained by IPTG induction of recombinant E. coli M15 cells containing the N-terminal region of LigB gee in PQE30 expression vector, was purified by Ni-NTA affinity chromatography and adsorbed on latex bead surface for performing latex agglutination test against Leptospirosis suspected wildlife field sera. A total of 80 wildlife sera samples were collec ted, including 27 wild feline sera samples (18 tigers, 8 lions, and 1 jaguar) obtained from Chhatbir zoo, Chandigarh, 42 sera samples (8 tigers, 4 lions and 6 leopards, 2 cheethals, 1 black buck, 12 buffaloes and 9 zoo staff) were received from Jodhpur zoo Rajasthan, 8 sera samples (4 tigers, 3 leopards, 1 lion) from Van Vihar National park, Bohpal, Madhya Pradesh and 3 sera samples (2 lions and 1 tiger) received from Biwani Mini zoo, Haryana, India. The result showed that sera were tested positive by rLigB based LAT, which were reconfirmed using microscopic agglutination test (MAT). The results from LAT were in concordance with MAT. In conclusion, rLigB based LAT is a rapid, pen site, reliable diagnostic tool of high sensitivity and specificity, under laboratory and field conditions, for the detection of Leptospirosis. Asian J. Med. Biol. Res. September 2020, 6(3): 440-448

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Conventional Rapid Latex Agglutination in Estimation of von Willebrand Factor: Method Revisited and Potential Clinical Applications

Journal of Immunology Research ◽

10.1155/2014/850810 ◽

2014 ◽

Vol 2014 ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Marianor Mahat ◽

Wan Zaidah Abdullah ◽

Che Maraina Che Hussin

Keyword(s):

Von Willebrand Factor ◽

Limit Of Detection ◽

Rapid Test ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Test Kit ◽

Agglutination Method ◽

Von Willebrand ◽

Willebrand Factor

Measurement of von Willebrand factor antigen (VWF : Ag) levels is usually performed in a specialised laboratory which limits its application in routine clinical practice. So far, no commercial rapid test kit is available for VWF : Ag estimation. This paper discusses the technical aspect of latex agglutination method which was established to suit the purpose of estimating von Willebrand factor (VWF) levels in the plasma sample. The latex agglutination test can be performed qualitatively and semiquantitatively. Reproducibility, stability, linearity, limit of detection, interference, and method comparison studies were conducted to evaluate the performance of this test. Semiquantitative latex agglutination test was strongly correlated with the reference immunoturbidimetric assay (Spearman’s rho = 0.946,P<0.001,n=132). A substantial agreement (κ=0.77) was found between qualitative latex agglutination test and the reference assay. Using the scoring system for the rapid latex test, no agglutination is with 0% VWF : Ag (control negative), 1+ reaction is equivalent to <20% VWF : Ag, and 4+ reaction indicates >150% VWF : Ag (when comparing with immunoturbidimetric assay). The findings from evaluation studies suggest that latex agglutination method is suitable to be used as a rapid test kit for the estimation of VWF : Ag levels in various clinical conditions associated with high levels and low levels of VWF : Ag.

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Comparative study of three tests (dye test, indirect haemagglutination test, latex agglutination test) for the detection of antibodies to Toxoplasma gondii in human sera.

Journal of Clinical Pathology ◽

10.1136/jcp.35.2.228 ◽

1982 ◽

Vol 35 (2) ◽

pp. 228-232 ◽

Cited By ~ 36

Author(s):

A H Balfour ◽

D G Fleck ◽

H P Hughes ◽

D Sharp

Keyword(s):

Toxoplasma Gondii ◽

Comparative Study ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Haemagglutination Test ◽

Indirect Haemagglutination ◽

Human Sera

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Effects of Pretreating Serum Samples on the Performance of a Latex Agglutination Test for Serodiagnosis of Paracoccidioidomycosis

Clinical and Vaccine Immunology ◽

10.1128/cvi.05274-11 ◽

2011 ◽

Vol 19 (3) ◽

pp. 386-390 ◽

Cited By ~ 5

Author(s):

Fabíola Silveira-Gomes ◽

Silvia Helena Marques-da-Silva

Keyword(s):

Sensitivity And Specificity ◽

Bacterial Infections ◽

Paracoccidioides Brasiliensis ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Serum Samples ◽

Content Type ◽

Human Sera ◽

Normal Human

ABSTRACTParacoccidioidomycosis (PCM) is a fungal disease caused byParacoccidioides brasiliensis, and Brazil is one of the principal countries where it is endemic. Diagnosis is based on the observation of buddingP. brasiliensisyeast in clinical specimens from patients; however, the sensitivity of the visualization of fungi is low, indicating that serological tests are used for early diagnosis. The double-immunodiffusion test (ID) is the “gold standard” test for serology in PCM, although the execution of this test requires the availability of laboratorial infrastructure. We report the improved performance of a latex agglutination test (LAT) by pretreating 30 serum samples from PCM patients and 71 controls (histoplasmosis and aspergillosis patients, patients with bacterial infections, and normal human sera) with a dilution buffer incubated at 37°C for 30 min. The sensitivity and specificity of the LAT test in the nonpretreated samples were 73% and 79%, respectively. However, when samples were pretreated, the sensitivity and specificity of the test increased to 90%. In this study, we did not observe cross-reactivity with histoplasmosis patient sera, but some reactions to sera from patients with aspergillosis and bacterial infections were noted. Normal human sera were not reactive in our tests. These results indicate the need for the elimination of heterologous reactions so that we can adequately use this method for screening cases of PCM.

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