Comparison between Microscopic Identification and Nested PCR for Detection of Cutaneous Leishmaniasis at Wasit Province

Cutaneous leishmaniasis (CL) is a widespread health problem and considered one of the endemic diseases in Iraq. The dermal lesion occurs due to an obligate intracellular Leishmania parasite, which transmits by the bite of the infected female sandfly. This study aims to identify Leishmania species in Thi-Qar province/South of Iraq and detect IL-17 level in serum of infected patients with L. tropica. The study was conducted in three local locations, Al-Hussein Teaching, Al-Suq Al-Shyokh General, and Al-Shatrah General Hospitals in the province for the period from the beginning of November 2018 to the end of October 2019. After clinical diagnosis, eighty out of two hundred forty-seven samples were selected for molecular examination by nested-PCR technique, where the lesion edge was injected by normal saline and pulled again to obtain the parasite DNA. Also, a measure of the IL-17 concentration level in serum of the patients with ELISA. The findings of the electrophoresis of the kinetoplast minicircle DNA gene showed that 65 samples were positive for cutaneous leishmaniasis, and observed two species of Leishmania spp. in the study area, 46 (57.5%) samples were L. tropica at 750 bp and 19 (23.75%) samples were L. major. Serum IL-17 concentration recorded a significant increase among patients infected with L. tropica at different infection stages than control samples. Generally, the Nested-PCR technique is an accurate method for diagnosing clinical samples and molecular determination of Leishmania parasites. L. tropica is the dominant specie that caused CL in Thi-Qar province, while L. major recorded a low incidence.

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Molecular characterization of Leishmania tropica and Leishmania major from stray dogs and patients in Saudi Arabia

10.21203/rs.2.22961/v2 ◽

2020 ◽

Author(s):

Abdullah D Alanazi ◽

Abdulazi S Alouffi S Alouffi ◽

Mohamed S Alyousif S Alyousif ◽

Abdulsadah A Rahi ◽

Magda A Ali ◽

...

Keyword(s):

Saudi Arabia ◽

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Leishmania Major ◽

Sand Fly ◽

Human Populations ◽

Leishmania Tropica ◽

Stray Dogs ◽

Wild Canids ◽

Control And Prevention

Abstract Background: Leishmania major and Leishmania tropica cause cutaneous leishmaniasis in humans and dogs in several parts of the world, with a large number of cases recorded in the Middle East. However, when occurring in sympatry, the role of each species of Leishmania in the epidemiology of cutaneous leishmaniasis (CL) is not clear. Methods: To determine the prevalence and to identify the species of Leishmania that infects humans and stray dogs in Riyadh and Al-Qassim (Saudi Arabia), 311 stray dogs and 27 human patients, suspected for Leishmania, were examined for CL by a nested PCR (nPCR).Results: Nested PCR (nPCR) detected seven patients (25.9%) positive for cutaneous leishmaniasis. Five patients from Riyadh were infected by L. major and two from Al-Qassim by L. tropica. In addition, five dogs (1.6%) were infected by L. tropica. Conclusions: This is one of the first molecular studies of leishmaniasis from Saudi Arabia. The relationship between the sand fly vectors and the reservoirs of both Leishmania spp. is still scarcely known in this region, and further epidemiological investigations of domestic and wild canids infected with L. major and L. tropica are needed towards a control and prevention of the infection in canine and human populations.

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Molecular characterization of Leishmania tropica and Leishmania major from stray dogs and patients in Saudi Arabia

10.21203/rs.2.22961/v1 ◽

2020 ◽

Author(s):

Abdullah D Alanazi ◽

Abdulazi S Alouffi S Alouffi ◽

Mohamed S Alyousif S Alyousif ◽

Abdulsadah A Rahi ◽

Magada A Ali ◽

...

Keyword(s):

Saudi Arabia ◽

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Leishmania Major ◽

Sand Fly ◽

Human Populations ◽

Leishmania Tropica ◽

Conventional Pcr ◽

Stray Dogs ◽

Control And Prevention

Abstract Background: Leishmania major and Leishmania tropica cause cutaneous leishmaniasis in humans and dogs in several parts of the world, with a large number of cases recorded in the Middle East. However, when occurring in sympatry in the same area, the role of each species in the epidemiology of cutaneous leishmaniasis is not clear.Methods: To determine the prevalence and to identify the species of Leishmania that infect humans and stray dogs in Riyadh and Al-Qaseem (Saudi Arabia), blood samples were collected from stray dogs (n = 311) and human patients (n = 27). A conventional PCR coupled with a nested PCR (n PCR) were applied in this study.Results: A conventional PCR coupled with a nested PCR (n PCR) showed that seven patients (25.9%) from Riyadh city and Al-Qaseem province were infected by L. major and L. tropica and five dogs (1.6%) by L. tropica.Conclusions: This is one of the first molecular studies of leishmaniasis from Saudi Arabia. The relationship between the sand fly vectors and the reservoirs of both Leishmania spp. is still scarcely known in this region, and further epidemiological investigations of domestic and wild canine infected with L. major and L. tropica are needed towards a control and prevention of the infection in canine and human populations.

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An Eco-Epidemiological Study on Zoonotic Cutaneous Leishmaniasis in Central Iran

Iranian Journal of Public Health ◽

10.18502/ijph.v50i2.5350 ◽

2021 ◽

Author(s):

Hossein NEZAMZADEH-EZHIYEH ◽

Hossein MIRHENDI ◽

Reza JAFARI ◽

Arshad VEYSI ◽

Yavar RASSI ◽

...

Keyword(s):

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Reservoir Host ◽

Leishmania Infection ◽

Sand Flies ◽

Wild Rodents ◽

Direct Examination ◽

North East ◽

Zoonotic Cutaneous Leishmaniasis ◽

Susceptibility Tests

Background: Leishmaniasis is an expanding neglected tropical disease in the world reporting from 98 countries including Iran. This study focused on eco-epidemiological determinants of the disease following a rapid and unexpected increase of leishmaniasis incidence in a strategic residential district in North-East of Isfahan County, Iran. Methods: This study was accomplished from Apr 2012 to Jan 2014 in a strategic residential zone in North-East of Isfahan County, Esfahan, Iran. Monthly activity, parity, Leishmania infection and susceptibility tests, were determined on sand flies. Some portion of inhabitants and school children were surveyed to find active or passive cases of leishmaniasis and also wild rodents were collected to determine reservoir host. Results: Totally 5223 sand flies belonging to Phlebotomus and Sergentomyia genus were collected and identified; Ph. papatasi was the dominant species and started to appear in May and disappeared in Oct. The majority of living dissected sand flies were unfed and parous. Ph. papatasi showed 4.6% Leishmania infection through direct examination and 39.54% by nested-PCR respectively. Phlebotomus papatasi was susceptible against deltametrin 0.05%. Totally 2149 people were surveyed and incidence and prevalence of zoonotic cutaneous leishmaniasis estimated as 45.39 and 314.40 per 1000 population. Rodents showed 73.91% and 80% Leishmania infection by direct examination and nested-PCR respectively. Conclusion: Cutaneous leishmaniasis due to L. major has been established in this area. Rodent control operation and personal protection are highly recommended to control the disease in this focus.

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PCR-positivity of gerbils and their ectoparasites for Leishmania spp. in a hyperendemic focus of zoonotic cutaneous leishmaniasis in central Iran

10.21203/rs.2.21837/v1 ◽

2020 ◽

Author(s):

Sahar Azarmi ◽

Alireza Zahraei-Ramazani ◽

Mehdi Mohebali ◽

Yavar Rassi ◽

Amir Ahmad Akhavan ◽

...

Keyword(s):

Cutaneous Leishmaniasis ◽

Infection Rate ◽

Microscopic Examination ◽

Nested Pcr ◽

Central Iran ◽

Nested Polymerase Chain Reaction ◽

Zoonotic Cutaneous Leishmaniasis ◽

Leishmania Spp ◽

Different Seasons

Abstract Background: Sand flies are the only known vectors of Leishmania parasites. Various arthropods, including ticks have been suggested as secondary vectors of Leishmania spp. many years ago. This study was conducted to determine PCR-positivity of zoonotic cutaneous leishmaniasis reservoir hosts and their ectoparasites for Leishmania spp. in central Iran. Methods: Microscopic examination and nested polymerase chain reaction (Nested-PCR) were used to detect and identify species of Leishmania and the results were confirmed by two methods, PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing. Results: Totally, 93 rodents (Rhomombis opimus (n=92) and Nesokia indica (n=1)) were captured during different seasons and 9 different species of ectoparasites were collected from them. Out of 92 collected R. opimus, 14 were positive for Leishmania spp. by microscopic examination while one N. indica and 77 R. opimus were positive by nested-PCR. The infection rate of rodents with Leishmania major and Leishmania turanica was 39.79% and 15.05%, respectively. Mixed natural infections with L. major and L. turanica were seen in 15.05% of rodents, in 7.53% of the rodents with L. major and Leishmania gerbilli, and in 6.45% of the rodents with the three species. The leishmanial infection rate of the rodents was the highest (94.29%) in summer and lowest (45.45%) in the winter. Moreover, 39 out of 54 fleas, 5 out of 8 mites, and 1 tick were PCR positive for Leishmania parasites. Conclusions: The highest rate of infection with L. major and L. turanica in R. opimus populations was observed in the summer and spring respectively, and the highest percentage of L. major and L. turanica coinfection was seen in the winter. It is suggested that the role of L. turanica and the probable role of ectoparasites in the transmission of epidemiology should be investigated carefully. Xenodiagnostic testing is recommended for future testing.

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PCR and Microscopic Identification of IsolatedLeishmania tropicafrom Clinical Samples of Cutaneous Leishmaniasis in Human Population of Kohat Region in Khyber Pakhtunkhwa

BioMed Research International ◽

10.1155/2014/861831 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 8

Author(s):

Nasser M. Abd El-Salam ◽

Sultan Ayaz ◽

Riaz Ullah

Keyword(s):

Cutaneous Leishmaniasis ◽

Rural Community ◽

Human Population ◽

Clinical Samples ◽

Leishmania Tropica ◽

Khyber Pakhtunkhwa ◽

Culture Techniques ◽

Microscopic Identification ◽

Khyber Pakhtunkhwa Province ◽

Very High

Leishmania tropicawas isolated from the clinical patients of cutaneous leishmaniasis in rural community of Kohat district in Khyber Pakhtunkhwa province and was identified through PCR, microscopy, and culture techniques. A total of 113 samples from the clinical patients were examined through PCR, microscopy, and culture which showed 87.61% (99/113), 53.98% (61/113), and 46.90% (53/113) prevalence. During the study, 186 bpLeishmania tropicawas identified through PCR. Thus the sensitivity of PCR is very high as compared to the conventional techniques.

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Nested PCR, RFLP and sequencing of nuclear ITS-rDNA genes detect Leishmania species in patients from an Iranian focus of zoonotic cutaneous leishmaniasis (ZCL)

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2011.08.1012 ◽

2011 ◽

Vol 44 (13) ◽

pp. S317

Author(s):

Ahmad Baghaei ◽

Parviz Parvizi ◽

Aref Amirkhani ◽

Alaee Novin Elnaz

Keyword(s):

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Leishmania Species ◽

Its Rdna ◽

Rdna Genes ◽

Zoonotic Cutaneous Leishmaniasis ◽

Pcr Rflp ◽

Nuclear Its

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Molecular characterization of Leishmania species from stray dogs and patients in Saudi Arabia

10.21203/rs.2.22961/v4 ◽

2020 ◽

Author(s):

Abdullah D Alanazi ◽

Abdulazi S Alouffi ◽

Mohamed S Alyousif ◽

Abdulsadah A Rahi ◽

Magda A Ali ◽

...

Keyword(s):

Saudi Arabia ◽

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Leishmania Major ◽

Leishmania Species ◽

Sand Fly ◽

Human Populations ◽

Stray Dogs ◽

Wild Canids ◽

Control And Prevention

Abstract Background: Leishmania major and Leishmania tropica cause cutaneous leishmaniasis in humans and dogs in several parts of the world, with a large number of cases recorded in the Middle East. However, when occurring in sympatry, the role of each species of Leishmania in the epidemiology of cutaneous leishmaniasis (CL) is not clear. Methods: To determine the prevalence and to identify the species of Leishmania that infects humans and stray dogs in Riyadh and Al-Qassim (Saudi Arabia), 311 stray dogs and 27 human patients, suspected for Leishmania, were examined for CL by a nested PCR (nPCR).Results: Nested PCR (nPCR) detected seven patients (25.9%) positive for cutaneous leishmaniasis. Five patients from Riyadh were infected by L. major and two from Al-Qassim by L. tropica. In addition, five dogs (1.6%) were infected by L. tropica. Conclusions: This is one of the first molecular studies of leishmaniasis from Saudi Arabia. The relationship between the sand fly vectors and the reservoirs of both Leishmania spp. is still scarcely known in this region, and further epidemiological investigations of domestic and wild canids infected with L. major and L. tropica are needed towards a control and prevention of the infection in canine and human populations.

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Molecular characterization of Leishmania species from stray dogs and patients in Saudi Arabia

10.21203/rs.2.22961/v5 ◽

2020 ◽

Author(s):

Abdullah D Alanazi ◽

Abdulazi S Alouffi ◽

Mohamed S Alyousif ◽

Abdulsadah A Rahi ◽

Magda A Ali ◽

...

Keyword(s):

Saudi Arabia ◽

Cutaneous Leishmaniasis ◽

Nested Pcr ◽

Leishmania Major ◽

Leishmania Species ◽

Sand Fly ◽

Human Populations ◽

Stray Dogs ◽

Wild Canids ◽

Control And Prevention

Abstract Background: Leishmania major and Leishmania tropica cause cutaneous leishmaniasis in humans and dogs in several parts of the world, with a large number of cases recorded in the Middle East. However, when occurring in sympatry, the role of each species of Leishmania in the epidemiology of cutaneous leishmaniasis (CL) is not clear. Methods: To determine the prevalence and to identify the species of Leishmania that infects humans and stray dogs in Riyadh and Al-Qassim (Saudi Arabia), 311 stray dogs and 27 human patients, suspected for Leishmania, were examined for CL by a nested PCR (nPCR).Results: Nested PCR (nPCR) detected seven patients (25.9%) positive for cutaneous leishmaniasis. Five patients from Riyadh were infected by L. major and two from Al-Qassim by L. tropica. In addition, five dogs (1.6%) were infected by L. tropica. Conclusions: This is one of the first molecular studies of leishmaniasis from Saudi Arabia. The relationship between the sand fly vectors and the reservoirs of both Leishmania spp. is still scarcely known in this region, and further epidemiological investigations of domestic and wild canids infected with L. major and L. tropica are needed towards a control and prevention of the infection in canine and human populations.

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