19. Waggoner JJ, Pinsky BA (2016) Molecular
diagnostics for human leptospirosis. Curr
Opin Infect Dis 29:440–445. https://doi.
org/10.1097/QCO.0000000000000295
20. Thaipadunpanit J, Chierakul W, Wuthiekanun
V et al (2011) Diagnostic accuracy of real-time
PCR assays targeting 16S rRNA and lipl32
genes for human leptospirosis in Thailand: a
case-control study. PLoS One 6:e16236.
https://doi.org/10.1371/journal.pone.
0016236
21. Agampodi S, Matthias M, Moreno A et al
(2012) Utility of quantitative polymerase
chain reaction in leptospirosis diagnosis: association of level of leptospiremia and clinical
manifestations in Sri Lanka. Clin Infect Dis
54:1249–1255.
https://doi.org/10.1093/
cid/cis035
22. Riediger IN, Stoddard RA, Ribeiro GS et al
(2017) Rapid, actionable diagnosis of urban
epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay. PLoS
Negl Trop Dis 11:e0005940. https://doi.org/
10.1371/journal.pntd.0005940
23. Mauk MG, Song J, Liu C et al (2018) Simple
approaches to minimally-instrumented, microfluidic-based point-of-care nucleic acid amplification tests. Biosensors (Basel) 8:17. https://
doi.org/10.3390/bios8010017
24. Sonthayanon P, Chierakul W, Wuthiekanun V
et al (2011) Accuracy of loop-mediated isothermal amplification for diagnosis of human
leptospirosis in Thailand. Am J Trop Med Hyg
84:614–620.
https://doi.org/10.4269/
ajtmh.2011.10-0473
25. Villumsen S, Pedersen R, Krogfelt KA et al
(2010) Expanding the diagnostic use of PCR
in leptospirosis: improved method for DNA
extraction from blood cultures. PLoS One 5:
e12095. https://doi.org/10.1371/journal.
pone.0012095
26. Dittrich S, Rudgard WE, Woods KL et al
(2016) The utility of blood culture fluid for
the molecular diagnosis of Leptospira: a prospective evaluation. Am J Trop Med Hyg
94:736–740.
https://doi.org/10.4269/
ajtmh.15-0674
27. Nhan TX, Teissier A, Roche C et al (2014)
Sensitivity of real-time PCR performed on
dried sera spotted on filter paper for diagnosis
of
leptospirosis.
J
Clin
Microbiol
52:3075–3077.
https://doi.org/10.1128/
JCM.00503-14
28. Matthias MA, Ricaldi JN, Cespedes M et al
(2008) Human leptospirosis caused by a new,
antigenically unique Leptospira associated with
a Rattus species reservoir in the Peruvian
Amazon. PLoS Negl Trop Dis 2:e213.
https://doi.org/10.1371/journal.pntd.
0000213
29. Slack AT, Kalambaheti T, Symonds ML et al
(2008) Leptospira wolffii sp. nov., isolated from
a human with suspected leptospirosis in
Thailand. Int J Syst Evol Microbiol
58:2305–2308. https://doi.org/10.1099/ijs.
0.64947-0
30. Chiriboga J, Barragan VA, Arroyo G et al
(2015) High prevalence of intermediate Leptospira spp. DNA in febrile humans from urban
and rural Ecuador. Emerg Infect Dis
21:2141–2147.
https://doi.org/10.3201/
eid2112.140659
31. Tsuboi M, Koizumi N, Hayakawa K et al
(2017) Imported Leptospira licerasiae infection
in traveler returning to Japan from Brazil.
Emerg Infect Dis 23:548–549. https://doi.
org/10.3201/eid2303.161262
32. Puche R, Ferre ´s I, Caraballo L et al (2018)
Leptospira venezuelensis sp. nov., a new member
of the intermediate group isolated from
rodents, cattle and humans. Int J Syst Evol
Microbiol 68:513–517. https://doi.org/10.
1099/ijsem.0.002528
33. Thibeaux R, Girault D, Bierque E et al (2018)
Biodiversity of environmental Leptospira:
improving identification and revisiting the
diagnosis. Front Microbiol 9:816. https://
doi.org/10.3389/fmicb.2018.00816
34. Ganoza CA, Matthias MA, Saito M et al (2010)
Asymptomatic renal colonization of humans in
the peruvian Amazon by Leptospira. PLoS Negl
Trop Dis 4:e612. https://doi.org/10.1371/
journal.pntd.0000612
35. Villumsen S, Pedersen R, Borre MB et al
(2012) Novel TaqMan
® PCR for detection of
Leptospira species in urine and blood: pit-falls
of in silico validation. J Microbiol Methods
91:184–190.
https://doi.org/10.1016/j.
mimet.2012.06.009
36. Woods K, Nic-Fhogartaigh C, Arnold C et al
(2018) A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with
fever in Laos. Clin Microbiol Infect 24:1017.
e1–1017.e7. https://doi.org/10.1016/j.cmi.
2017.10.017
37. Backstedt BT, Buyuktanir O, Lindow J et al
(2015) Efficient detection of pathogenic leptospires using 16S ribosomal RNA. PLoS One
10:e0128913. https://doi.org/10.1371/jour
nal.pone.0128913
38. Waggoner JJ, Balassiano I, Mohamed-Hadley
A et al (2015) Reverse-transcriptase PCR
detection of Leptospira: absence of agreement
Laboratory Diagnosis of Leptospirosis
285
diagnostics for human leptospirosis. Curr
Opin Infect Dis 29:440–445. https://doi.
org/10.1097/QCO.0000000000000295
20. Thaipadunpanit J, Chierakul W, Wuthiekanun
V et al (2011) Diagnostic accuracy of real-time
PCR assays targeting 16S rRNA and lipl32
genes for human leptospirosis in Thailand: a
case-control study. PLoS One 6:e16236.
https://doi.org/10.1371/journal.pone.
0016236
21. Agampodi S, Matthias M, Moreno A et al
(2012) Utility of quantitative polymerase
chain reaction in leptospirosis diagnosis: association of level of leptospiremia and clinical
manifestations in Sri Lanka. Clin Infect Dis
54:1249–1255.
https://doi.org/10.1093/
cid/cis035
22. Riediger IN, Stoddard RA, Ribeiro GS et al
(2017) Rapid, actionable diagnosis of urban
epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay. PLoS
Negl Trop Dis 11:e0005940. https://doi.org/
10.1371/journal.pntd.0005940
23. Mauk MG, Song J, Liu C et al (2018) Simple
approaches to minimally-instrumented, microfluidic-based point-of-care nucleic acid amplification tests. Biosensors (Basel) 8:17. https://
doi.org/10.3390/bios8010017
24. Sonthayanon P, Chierakul W, Wuthiekanun V
et al (2011) Accuracy of loop-mediated isothermal amplification for diagnosis of human
leptospirosis in Thailand. Am J Trop Med Hyg
84:614–620.
https://doi.org/10.4269/
ajtmh.2011.10-0473
25. Villumsen S, Pedersen R, Krogfelt KA et al
(2010) Expanding the diagnostic use of PCR
in leptospirosis: improved method for DNA
extraction from blood cultures. PLoS One 5:
e12095. https://doi.org/10.1371/journal.
pone.0012095
26. Dittrich S, Rudgard WE, Woods KL et al
(2016) The utility of blood culture fluid for
the molecular diagnosis of Leptospira: a prospective evaluation. Am J Trop Med Hyg
94:736–740.
https://doi.org/10.4269/
ajtmh.15-0674
27. Nhan TX, Teissier A, Roche C et al (2014)
Sensitivity of real-time PCR performed on
dried sera spotted on filter paper for diagnosis
of
leptospirosis.
J
Clin
Microbiol
52:3075–3077.
https://doi.org/10.1128/
JCM.00503-14
28. Matthias MA, Ricaldi JN, Cespedes M et al
(2008) Human leptospirosis caused by a new,
antigenically unique Leptospira associated with
a Rattus species reservoir in the Peruvian
Amazon. PLoS Negl Trop Dis 2:e213.
https://doi.org/10.1371/journal.pntd.
0000213
29. Slack AT, Kalambaheti T, Symonds ML et al
(2008) Leptospira wolffii sp. nov., isolated from
a human with suspected leptospirosis in
Thailand. Int J Syst Evol Microbiol
58:2305–2308. https://doi.org/10.1099/ijs.
0.64947-0
30. Chiriboga J, Barragan VA, Arroyo G et al
(2015) High prevalence of intermediate Leptospira spp. DNA in febrile humans from urban
and rural Ecuador. Emerg Infect Dis
21:2141–2147.
https://doi.org/10.3201/
eid2112.140659
31. Tsuboi M, Koizumi N, Hayakawa K et al
(2017) Imported Leptospira licerasiae infection
in traveler returning to Japan from Brazil.
Emerg Infect Dis 23:548–549. https://doi.
org/10.3201/eid2303.161262
32. Puche R, Ferre ´s I, Caraballo L et al (2018)
Leptospira venezuelensis sp. nov., a new member
of the intermediate group isolated from
rodents, cattle and humans. Int J Syst Evol
Microbiol 68:513–517. https://doi.org/10.
1099/ijsem.0.002528
33. Thibeaux R, Girault D, Bierque E et al (2018)
Biodiversity of environmental Leptospira:
improving identification and revisiting the
diagnosis. Front Microbiol 9:816. https://
doi.org/10.3389/fmicb.2018.00816
34. Ganoza CA, Matthias MA, Saito M et al (2010)
Asymptomatic renal colonization of humans in
the peruvian Amazon by Leptospira. PLoS Negl
Trop Dis 4:e612. https://doi.org/10.1371/
journal.pntd.0000612
35. Villumsen S, Pedersen R, Borre MB et al
(2012) Novel TaqMan
® PCR for detection of
Leptospira species in urine and blood: pit-falls
of in silico validation. J Microbiol Methods
91:184–190.
https://doi.org/10.1016/j.
mimet.2012.06.009
36. Woods K, Nic-Fhogartaigh C, Arnold C et al
(2018) A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with
fever in Laos. Clin Microbiol Infect 24:1017.
e1–1017.e7. https://doi.org/10.1016/j.cmi.
2017.10.017
37. Backstedt BT, Buyuktanir O, Lindow J et al
(2015) Efficient detection of pathogenic leptospires using 16S ribosomal RNA. PLoS One
10:e0128913. https://doi.org/10.1371/jour
nal.pone.0128913
38. Waggoner JJ, Balassiano I, Mohamed-Hadley
A et al (2015) Reverse-transcriptase PCR
detection of Leptospira: absence of agreement
Laboratory Diagnosis of Leptospirosis
285