Comparison of the Polymerase Chain Reaction and Serology for the Detection of Canine Visceral Leishmaniasis

Ashford, David A.; Degrave, Wim; Eulalio, Conceicao; Badaró, Roberto; Lopes, Ulisses Gazos; Miranda, José Carlos; David, John R.; Sherlock, Ítalo Rodrigues de Araújo; Bozza, Marcelo T.; Barker, Robert H.; Freire, Maria do Carmo Matias; Fernandes, Octávio

doi:10.4269/ajtmh.1995.53.251

Cited by 116 publications

(80 citation statements)

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“…It consisted of an amplification of the acidic ribosomal phospohoprotein fragment of houskeeping canine gene with PO3 and PO5 primers amplifying a 469 pb PCR product (Ashford et al, 1995). If the inhibition control was negative, a five-fold dilution of the DNA was applied for the subsequent PCR.…”

Section: Pcrmentioning

confidence: 99%

“…Several studies suggesting that the rate of infection is higher than the figures found by serological investigations (Lachaud et al, 2002;Leontides et al, 2002;Oliva et al, 2006). Several molecular methods had been developed for the detection of Leishmania (Van Eys et al, 1992;Ashford et al, 1995;Spanakos et al, 2002). Though PCR was already used in the diagnosis of human leishmaniasis in Tunisia (Chargui et al, 2005), this is the first study to investigate the prevalence of CanL with molecular methods compared to serological and parasitological ones.…”

mentioning

confidence: 99%

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Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

et al. 2009

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Summary :The aim of this study was to assess the use of parasitological, serological and molecular methods for the detection of Leishmania infection in blood of 67 dogs and to investigate the prevalence of canine leishmaniasis (CanL) in Kairouan (central Tunisia), an area known to be of reduced endemicity and has not been studied since 1973. Veterinarians clinically examined all dogs, and the titer of anti-Leishmania antibodies was determined by indirect immune-fluorescence antibody test. The presence of Leishmania was performed by PCR and in vitro culture. IFAT was positive in 12 % of dogs and promastigote form of the parasite was isolated by in vitro culture from only 4.5 % of them. However, DNA of Leishmania was detected by PCR in 20.9 % of dogs. PCR was more sensitive than IFAT (p = 0.004) and in vitro culture (p < 10 -5 ). A prevalence of 21 % was found in Kairouan, which is significant high (p < 10 -3 ) when compared to that of thirty years ago. This state is in correlation with the increase in other Mediterranean countries. Furthermore, 50 % of positive dogs were asymptomatic. Preventive measures must be taken against these dogs as for symptomatic ones since their role in the transmission of the infection to vectors has been proven.

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Section: Pcrmentioning

confidence: 99%

mentioning

confidence: 99%

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

et al. 2009

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“…The polymerase chain reaction (PCR) represents a new option among them (2)(3)(4)(5). PCR has been shown to be highly sensitive for the diagnosis of ACL, with rapid detection of leishmania (6)(7)(8)(9)(10)(11). PCR can also be used for the typing of leishmania isolated from clinical material, insect vectors, reservoirs, or culture media (12,13).…”

Section: Introductionmentioning

confidence: 99%

Comparison of the specificity of PCR and the histopathological detection of leishmania for the diagnosis of American cutaneous leishmaniasis

Medeiros

Rodrigues

Roselino

2002

Braz J Med Biol Res

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More precise and rapid diagnostic methods for American cutaneous leishmaniasis (ACL) are necessary because of the growing number of cases observed in Brazil, including the northeastern region of the State of São Paulo. We applied PCR to 54 skin or mucosal biopsies from patients with a clinical and/or laboratory diagnosis of ACL using primers 13A and 13B, with positive results being obtained for 82% of the samples. When the PCR results were compared to those of histopathological leishmania detection, PCR showed superior results with 81.5% sensitivity and 95% CI of 68.0-95.1%. The Montenegro skin test (MST) was positive in 88.7% of patients. Since MST cannot be used as a diagnostic tool in endemic areas, the present results strongly suggest the use of PCR for the etiological confirmation of ACL, with emphasis on the mucosal form. Correspondence

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“…This same author has reported that given the high sensitivity of the test, it is possible to perform PCR with just 10 parasites and to detect the band in the electrophoresis gel. Nevertheless, PCR does not always detect the parasite, since inhibition of the reaction may occur in some types of sample 3 , presenting a decrease in the sensitivity of the test to 60%-70%. Nevertheless, this is one of the most promising techniques for the identification of the parasite, given the fact that it is noninvasive and the circulating parasites can be detected in a sample of blood or leukocyte cream.…”

Section: Discussionmentioning

confidence: 99%

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Nogueira

Galati

2006

Rev. Inst. Med. trop. S. Paulo

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SUMMARYLeishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µL and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume.The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time.These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.

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Comparison of the Polymerase Chain Reaction and Serology for the Detection of Canine Visceral Leishmaniasis

Cited by 116 publications

References 0 publications

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

Comparison of the specificity of PCR and the histopathological detection of leishmania for the diagnosis of American cutaneous leishmaniasis

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

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