Recombinant LipL32 Protein for Leptospirosis Detection in Indonesia

Sumarningsih,; Tarigan, Simson; Susanti, _; Kusmiyati,

doi:10.1016/j.proche.2016.01.004

Cited by 9 publications

(11 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Antigen-antibody binding was also found in serum from bovine immunized with rLipL32 (lane 6) but preimmune bovine serum could not response to LipL32 protein. These results demonstrated that the rLipL32 could induce high antibody response in rabbit and bovine, indicating the high antigenicity and immunogenicity of protein LipL32, as reported from the preliminary study which showed that rLipL32 protein could detect serum from bovine positive MAT (Sumarningsih et al 2016). …”

Section: Western Blotting Of Anti Rlipl32 Serumsupporting

confidence: 71%

“…Studies conducted previously by Bomfim et al (2005) and Dey et al (2004) reported high sensitivity and accuracy for ELISA LipL32 in bovine and canine, respectively, compared to MAT. However, in our preliminary study using 13 serum sample showed inconsistent results between MAT and ELISA LipL32 for detection of Leptospira in bovine (Sumarningsih et al 2016). The inconsistent results of the previous study were: some positive sera on MAT showed negative responsense on ELISA LipL32 and some negative sera on MAT were found to be positive on ELISA LipL32.…”

Section: Introductioncontrasting

confidence: 56%

“…Bovine anti LipL32 serum was collected for LipL32 titre analysis. Similar procedure for MAT was conducted as described by Sumarningsih et al (2016) to determine the titre of all sera collected in this study.…”

Section: Production Of Antibody To Rlipl32 Proteinmentioning

confidence: 99%

See 2 more Smart Citations

Preliminary Study: Characterisation of Antibody for rLipl32 Protein of Leptospira

Sumarningsih¹,

Tarigan²

2016

Proceedings of International Seminar on Livestock Production and Veterinary Technology

Self Cite

View full text Add to dashboard Cite

Leptospirosis infection is commonly determined by positive response antibody in microaglutination test (MAT). The test requires panels of Leptospira from complete serovars and uses infectious Leptospira, which is subcultured weekly. Accordingly, other alternative serological tests are developed for MAT. One of them is ELISA using recombinant LipL32 protein (rLipL32). High sensitivity and specificity of ELISA rLipL32 for Leptospirosis detection in bovine and canine has been reported previously. In this study, we produced antibody to rLipL32 protein with high titre. Results on western blotting and ELISA showed anti-LipL32 antibody was highly binding to rLipL32 protein. In contrast, MAT results for LipL32 antibody was found to be negative response, indicating that titre antibody to LipL32 does not positively correlate to MAT.

show abstract

Section: Western Blotting Of Anti Rlipl32 Serumsupporting

confidence: 71%

Section: Introductioncontrasting

confidence: 56%

See 1 more Smart Citation

Preliminary Study: Characterisation of Antibody for rLipl32 Protein of Leptospira

Sumarningsih¹,

Tarigan²

2016

Proceedings of International Seminar on Livestock Production and Veterinary Technology

Self Cite

View full text Add to dashboard Cite

show abstract

“…The plasmid of pRSET-C-LipL32 was purified from E.coli BL21 cells and sent to 1 st base (http://www.baseasia.com/dna_sequencing/) for sequencing as described in previous study (Sumarningsih et al 2016). Amino acid sequences of LipL32 from Leptospira serovar Pomona, Hardjo and Grippotyphosa were obtained from GeneBank with accession number AY609326, AY442332 and AY609327, respectively.…”

Section: Sequence Analysismentioning

confidence: 99%

“…MAT for bovine serum was performed as described in previous study (Sumarningsih et al 2016). Briefly, panels of antigens were added into serum samples that had been diluted in PBS, pH 7.4 at 1 : 50, 1 : 100, 1 : 400, 1 : 1600.…”

Section: Micro Agglutination Test (Mat)mentioning

confidence: 99%

Evaluation of LipL32 ELISA for detection of bovine leptospirosis in West Java

Sumarningsih¹,

Susanti²,

Tarigan³

2018

JITV

Self Cite

View full text Add to dashboard Cite

The current diagnosis of leptospirosis, micro Agglutination Test (MAT) and isolation, is expensive, impractical and technically demanding. This study was aimed at developing an ELISA based on recombinant LipL32 as a practical, inexpensive test for Leptospirosis. The DNA encoding LipL32 was isolated from Leptospira pomona, inserted into pRSET-C plasmid then expressed in E.coli BL21 as a poly-histidine-tagged protein. The amount of LipL32 protein, which was purified from the supernatant of lysed cells by a Ni-NTA column, was 1mg/l culture. This purified LipL32 was used as the coating antigen at 5µg/ml. The accuracy of ELISA was evaluated based on ROC analysis, by comparing the ELISA and MAT results of 517 bovine sera. Result in this study showed that the area under curve (AUC) was 0.853, which categorised the LipL32 ELISA as a “moderately accurate” test and indicates that the ELISA was able to differentiate positive and negative Leptospirosis serum. The result also showed ELISA LipL32 could detect serum positive MAT to Hardjo, Grippotyphosa, Tarrasovi, Rachmati and Bataviae. The optimal cut off for OD ELISA determined based on ROC curve was 0.504, and it showed sensitivity and specificity of ELISA LipL32 relative to MAT were 86.0% and 69.5%, respectively. Overall, the result in this study showed that ELISA LipL32 can be used as a rapid test for identification of anti-Leptospira antibodies in bovine.

show abstract