Combined antibody and DNA detection for early diagnosis of leptospirosis after a disaster

Iwasaki, H.; Chagan-Yasutan, Haorile; Leano, Prisca Susan A.; Koizumi, Nobuo; Nakajima, Chie; Taurustiati, Delsi; Hanan, Firmanto; Lacuesta, Talitha Lea; Ashino, Yugo; Suzuki, Yasuhiko; Gloriani, Nina G.; Telan, Elizabeth Freda O.; Hattori, Toshio

doi:10.1016/j.diagmicrobio.2016.01.001

Cited by 20 publications

(17 citation statements)

References 38 publications

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“…We therefore suggest that the urinary markers examined here reflect different types of damage caused by kidney failure resulting from Leptospira infection; uDA1 may reflect the involvement of neutrophils during kidney injury, whereas uNGAL and uNAG reflect both neutrophil involvement and tubular dysfunction. Our previous data showed the presence of Leptospira DNA in the urine of some of the patients [13], and uDA1 was also positive in kidney derived-DNA-positive cases (unpublished observation), further supporting that neutrophil activation is caused by bacteria. Very recently, it was shown that the leptospiral outer membrane protein LipL32 induces inflammation, including leukocyte infiltration and kidney injury, in zebrafish larvae [36].…”

Section: Discussionsupporting

confidence: 53%

“…A retrospective study was performed in 135 patients admitted to San Lazaro Hospital (Manila, Philippines) in 2012 with clinical suspicion of leptospirosis; 112 of them were confirmed positive with one of following diagnostic tools using serum or urine samples: the microscopic agglutination test [37], immunochromatographic assay (Standard Diagnostics, Yongin, Korea), enzyme-linked immunosorbent assay (ELISA; Diagnostic Automation, Calabasas, CA, USA), loop-mediated isothermal amplification, and real-time PCR [13,38]. The remaining 23 patients were confirmed to have non-leptospirosis illnesses of unknown origin and served as controls.…”

Section: Methodsmentioning

confidence: 99%

“…Azithromycin was also shown to inhibit neutrophil activation [10,11], and involvement of neutrophils and elevation of neutrophil-related activation markers such as neutrophil gelatinase-associated lipocalin (NGAL) have been reported in leptospirosis patients [12]. We recently demonstrated that urinary leptospiral DNA, some of which is presumably from phagocytized bacteria in leukocytes, increased the detection rate of leptospirosis [13]. Therefore, we tried to identify urinary markers specific to neutrophil involvement in this disease in addition to NGAL.…”

Section: Introductionmentioning

confidence: 99%

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Urine Levels of Defensin α1 Reflect Kidney Injury in Leptospirosis Patients

Chagan-Yasutan

Chen

Lacuesta³

et al. 2016

IJMS

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Leptospirosis is a zoonotic disease whose severe forms are often accompanied by kidney dysfunction. In the present study, urinary markers were studied for potential prediction of disease severity. Urine samples from 135 patients with or without leptospirosis at San Lazaro Hospital, the Philippines, were analyzed. Urine levels of defensin α1 (uDA1) were compared with those of neutrophil gelatinase-associated lipocalin (uNGAL) and N-acetyl-β-d-glucosidase (uNAG). Serum creatinine (Cr) was used as a marker of kidney injury. The levels of uDA1/Cr, uNGAL/Cr, and uNAG/Cr were positive in 46%, 90%, and 80% of leptospirosis patients, and 69%, 70%, and 70% of non-leptospirosis patients, respectively. In leptospirosis patients, the correlation of uDA1/Cr, uNGAL/Cr and uNAG/Cr levels with serum Cr were r = 0.3 (p < 0.01), r = 0.29 (p < 0.01), and r = 0.02 (p = 0.81), respectively. uDA1/Cr levels were correlated with uNGAL/Cr levels (r = 0.49, p < 0.01) and uNAG/Cr levels (r = 0.47, p < 0.0001) in leptospirosis patients. These findings suggest that uDA1, uNGAL, and uNAG were elevated in leptospirosis patients and reflected various types of kidney damage. uDA1 and uNGAL can be used to track kidney injury in leptospirosis patients because of their correlation with the serum Cr level.

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Section: Discussionsupporting

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Urine Levels of Defensin α1 Reflect Kidney Injury in Leptospirosis Patients

Chagan-Yasutan

Chen

Lacuesta³

et al. 2016

IJMS

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“…Although the timing of Leptospira excretion in urine in humans is not clearly defined, our data support the findings of Iwasaki et al. [29] that Leptospira DNA detection by qPCR in urine occurs both early and late in the acute phase of leptospirosis.…”

Section: Discussionmentioning

confidence: 98%

A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

Woods

NicFhogartaigh

Arnold

et al. 2018

Clinical Microbiology and Infection

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ObjectivesTo compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos.MethodsSerum, buffy coat and urine samples were collected on admission, and follow-up serum ∼10 days later. Leptospira spp. culture and microscopic agglutination tests (MAT) were performed as reference standards. Bayesian latent class modelling was performed to estimate sensitivity and specificity of each diagnostic test.ResultsIn all, 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) were MAT positive, 76/787 (9.7%) were rrs qPCR positive and 20/787 (2.5%) were 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in at least one sample. Estimated sensitivity and specificity (with 95% CI) of 16SrRNA/LipL32 qPCR on serum (53.9% (33.3%–81.8%); 99.6% (99.2%–100%)), buffy coat (58.8% (34.4%–90.9%); 99.9% (99.6%–100%)) and urine samples (45.0% (27.0%–66.7%); 99.6% (99.3%–100%)) were comparable with those of rrs qPCR, except specificity of 16SrRNA/LipL32 qPCR on urine samples was significantly higher (99.6% (99.3%–100%) vs. 92.5% (92.3%–92.8%), p <0.001). Sensitivities of MAT (16% (95% CI 6.3%–29.4%)) and culture (25% (95% CI 13.3%–44.4%)) were low. Mean positive Cq values showed that buffy coat samples were more frequently inhibitory to qPCR than either serum or urine (p <0.001).ConclusionsSerum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. Quantitative PCR on admission is a reliable rapid diagnostic tool, performing better than MAT or culture, with significant implications for clinical and epidemiological investigations of this global neglected disease.

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“…The combination of DNA detection and antibody detection will enhance the accuracy for diagnosis of leptospirosis (Iwasaki et al 2016). …”

Section: Loop-mediated Isothermal Amplification Assay As Point-ofcarementioning

confidence: 99%

Development of Point-of-Care Testing for Disaster-Related Infectious Diseases

Hattori

Chagan-Yasutan

Shiratori

et al. 2016

Tohoku J. Exp. Med.

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After disaster, the victims lose their safe lives and are even exposed to nature where they could suffer from animal bites and vectors followed by suffering from zoonosis or vector-born diseases. Because of the urgent need for rapid and cheap diagnosis for infectious diseases after disaster, anonymous questionnaire clarified that leptospirosis, dengue, diarrhea, and cholera were recognized as common disaster-related infections in the Philippines, while diarrhea and pneumonia were more common in Indonesia. It should also be noted that infectious disease itself such as tuberculosis associated with acquired immune deficiency syndrome in South Africa is a disaster. Thus, the possible occurrence of similar situation in Asia should be prevented. We have conducted an international collaborative research in the Philippines and Indonesia on dengue virus, leptospira and mycobacterium tuberculosis (MTB) infectious diseases. Development of point-of-care testing for molecular diagnosis and disease severity was the principal purpose of the research. Loop-mediated isothermal amplification assay, which does not require a source of electricity, was developed for leptospirosis, dengue and MTB and has been proved to be useful where resource is limited. The plasma levels of matricellular proteins, including galectin-9 and osteopontin, were found to reflect the disease severities in dengue virus and MTB infection, probably because matricellular proteins are one of the most functional extracellular proteins that are associated with inflammatory edema. The study on disaster-related infectious disease facilitates the international cooperation for development of point-of-care testing for tropical infectious diseases.

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Combined antibody and DNA detection for early diagnosis of leptospirosis after a disaster

Cited by 20 publications

References 38 publications

Urine Levels of Defensin α1 Reflect Kidney Injury in Leptospirosis Patients

Urine Levels of Defensin α1 Reflect Kidney Injury in Leptospirosis Patients

A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

Development of Point-of-Care Testing for Disaster-Related Infectious Diseases

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