Polymerase chain reaction and loop-mediated isothermal amplification targeting lic13162, lic20239, and lipL32 genes for leptospirosis diagnosis

Pacce, Violetta Dias; Souza, Margarida Neves; Oliveira, Natasha Rodrigues de; Kremer, Frederico Schmitt; Jorge, Sérgio; Ikuta, Nilo; Lunge, Vagner Ricardo; Dellagostin, Odir Antônio

doi:10.1007/s42770-022-00698-1

Cited by 2 publications

(2 citation statements)

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“…An early-stage detection of LipL32 by an integrated lateral-flow immunoassay with an electrochemical readout showed a superior performance. It is reported to be comparable with the real-time PCR technique [70 ▪ ]. PCR and loop-mediated isothermal amplification targeting of lipL32 could detect pathogenic leptospira directly from clinical samples.…”

Section: Challenges In the Laboratory Work Upmentioning

confidence: 89%

“…PCR and loop-mediated isothermal amplification targeting of lipL32 could detect pathogenic leptospira directly from clinical samples. Diagnostic performance of lipL32-LAMP presented 84.2% sensitivity and 93.2% specificity [70 ▪ ]. The recombinase polymerase amplification-CRISPR/Cas12 (clustered regularly-interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 12a) targeting the lipL32 gene demonstrated good sensitivity and excellent specificity for detection of leptospirosis [71].…”

Section: Challenges In the Laboratory Work Upmentioning

confidence: 99%

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Ocular leptospirosis: lack of awareness among ophthalmologists and challenges in diagnosis

Sivakumar¹

2022

Current Opinion in Ophthalmology

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Purpose of reviewSystemic leptospirosis exists worldwide. But leptospiral uveitis, an important late complication is not identified by ophthalmologists in several countries.Recent findingsIn the last 18 months numerous publications, especially on epidemiology, surveillances, and novel rapid diagnostic kits to test at the point of care site on leptospirosis have been published from all over the world. However, publications from ophthalmologists are very scarce. Remarkably ophthalmologists should know the global burden of leptospirosis, prevalence of the disease in their country, demographic factors associated, risk factors, and systemic signs to elicit relevant history and travel history. They should be aware of recent advances in investigations to confirm their clinical diagnosis.SummaryIt is quite evident from this work that leptospirosis is prevalent worldwide. Ophthalmologists’ awareness has to improve to identify the etiological diagnosis. They should have access to simple, less expensive, and less cumbersome laboratory tests.

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Section: Challenges In the Laboratory Work Upmentioning

confidence: 89%

Section: Challenges In the Laboratory Work Upmentioning

confidence: 99%

Ocular leptospirosis: lack of awareness among ophthalmologists and challenges in diagnosis

Sivakumar¹

2022

Current Opinion in Ophthalmology

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Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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Background and Aim: Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis. Materials and Methods: Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting. Results: PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus. Conclusion: IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis. Keywords: anti-rLipL32 serum, immunoglobulin G anti-rLipL32 antibody, Leptospira, rLipL32 protein.

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Polymerase chain reaction and loop-mediated isothermal amplification targeting lic13162, lic20239, and lipL32 genes for leptospirosis diagnosis

Cited by 2 publications

References 50 publications

Ocular leptospirosis: lack of awareness among ophthalmologists and challenges in diagnosis

Ocular leptospirosis: lack of awareness among ophthalmologists and challenges in diagnosis

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

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