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

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

Asian Journal of Medical and Biological Research ◽

10.3329/ajmbr.v6i2.48054 ◽

2020 ◽

Vol 6 (2) ◽

pp. 229-236

Author(s):

Yosef Deneke ◽

Rajib Deb ◽

SM Lutful Kabir

Keyword(s):

Acute Infection ◽

Latex Agglutination ◽

Agglutination Test ◽

The Body ◽

Latex Agglutination Test ◽

City Hospital ◽

Indian Veterinary Research Institute ◽

Test Kit ◽

Human Sera ◽

Sera Samples

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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Expression of Recombinant Leptospiral Surface Lipoprotein-Lsa27 in E. coli and Its Evaluation for Serodiagnosis of Bovine Leptospirosis by Latex Agglutination Test

Molecular Biotechnology ◽

10.1007/s12033-020-00278-4 ◽

2020 ◽

Vol 62 (11-12) ◽

pp. 598-610

Author(s):

Anusha Alamuri ◽

K. Vinod Kumar ◽

S. SowjanyaKumari ◽

L. Linshamol ◽

R. Sridevi ◽

...

Keyword(s):

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

E Coli ◽

Bovine Leptospirosis

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Performance Comparison of a fliCh7 Real-Time PCR Assay with an H7 Latex Agglutination Test for Confirmation of the H Type of Escherichia coli O157:H7†

Journal of Food Protection ◽

10.4315/0362-028x-72.10.2195 ◽

2009 ◽

Vol 72 (10) ◽

pp. 2195-2197 ◽

Cited By ~ 7

Author(s):

NEELAM NARANG ◽

PINA M. FRATAMICO ◽

GLENN TILLMAN ◽

KITTY PUPEDIS ◽

WILLIAM C. CRAY

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Escherichia Coli O157 ◽

Pcr Assay ◽

E Coli ◽

Reference Strains ◽

Pcr Test

Escherichia coli O157:H7 is a foodborne pathogen that causes hemorrhagic colitis and hemolytic uremic syndrome. Positive identification of E. coli O157:H7 is made using biochemical tests and specific antisera or latex agglutination reagents for the O157 and H7 antigens. However, under certain conditions, some E. coli O157:H7 isolates can appear to be nonreactive with H7 antisera and may require multiple passages on motility medium to restore H7 antigenicity. In this study, we compared the performance of a real-time PCR test with that of a method using latex agglutination reagents to detect the presence of the fliCh7 gene or the H7 antigen, respectively, in E. coli O157:H7 isolates. One hundred twenty-six E. coli strains were tested including reference strains and strains isolated from meat. Lyophilized E. coli O157:H7 isolates were rehydrated and were plated on sheep blood agar without passage on motility medium. All strains were analyzed in parallel by a real-time PCR test targeting the fliCh7 gene and by a latex agglutination test that detects the H7 antigen. The real-time PCR assay showed 100% agreement with the H7 status reported for reference strains and E. coli O157:H7 meat isolates. The latex agglutination test results agreed with the H7 status reported for the E. coli O157:H7 reference strains and non-O157:H7 strains, except for one, E. coli O117:H7; however, 42% (42 of 100) of the E. coli O157:H7 meat isolates tested negative for the H7 antigen by latex agglutination. The real-time fliCh7 PCR test can be used to confirm E. coli O157:H7 strains that are not expressing the immunoreactive H7 antigen.

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Development of a Latex Agglutination Test for Detecting Pathogenic Burkholderia and its Approbation in the Endemic Regions of Vietnam

Problems of Particularly Dangerous Infections ◽

10.21055/0370-1069-2020-4-133-138 ◽

2021 ◽

pp. 133-138

Author(s):

D. M. Frolov ◽

N. N. Teteryatnikova ◽

T. L.A. Bui ◽

I. B. Zakharova ◽

N. P. Khrapova

Keyword(s):

Burkholderia Cepacia ◽

High Sensitivity ◽

Polymyxin B ◽

Latex Agglutination ◽

Agglutination Test ◽

Latex Agglutination Test ◽

Antigenic Structure ◽

Bacterial Cultures ◽

Freeze Dried ◽

Latex Test

The aim of the work was development of a monoclonal antibody-based latex agglutination test to identify the causative agent of melioidosis, and the approbation of a freeze-dried experimental preparation for screening of environmental bacterial isolates in Vietnam.Materials and methods. The carriers of specific antibodies were polyacrolein latex particles with active aldehyde groups on the surface. Typical strains of the causative agents of melioidosis and glanders with a full-fledged antigenic structure, as well as the strains Burkholderia thailandensis, Burkholderia cepacia, Pseudomonas aeruginosa, and Pseudomonas putida were used to control the test specificity. The latex agglutination reaction was carried out on plastic Petri dishes with daily bacterial cultures, from which suspensions were prepared at a concentration of 1–2·109 m.c./ml. The results of the reaction were registered visually for 5–8 min using a 4-cross system against a dark background under lighting. The reaction to 3–4 crosses was recorded as positive. Colonies suspected of belonging to pathogenic Burkholderia from primary inoculations were transferred to L-agar with polymyxin B and grown for 36 hours at (37±1) °C. The species of the selected suspicious colonies was determined by multiplex PCR.Results and discussion. With collection strains, latex test demonstrated high sensitivity agglutinating 97.7 % of B. pseudomallei and all B. mallei strains. At the same time, it was negative with B. thailandensis, B. cepacia, P. aeruginos and P. putida. In microbiological screening of bacterial cultures isolated from environmental objects, the latex test had a diagnostic sensitivity of 89.4 %. Using the latex test at the stage of primary screening, it is possible to significantly reduce the time when processing a lot of samples received for analysis, as well as to reduce the consumption of reagents used at the subsequent stages of identification.

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