scholarly journals Detection of Brucella sp. and Leptospira sp. in dogs using conventional polymerase chain reaction

2014 ◽  
Vol 58 (4) ◽  
pp. 527-531
Author(s):  
Faham Khamesipour ◽  
Abbas Doosti ◽  
Mohsen Fard Emadi ◽  
Babafela Awosile
Keyword(s):  
Polymerase Chain Reaction ◽  
Age Groups ◽  
Conventional Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Blood Samples ◽  
Stray Dogs ◽  
Supplementary Method ◽  
Significant Difference ◽  
Polymerase Chain ◽  
First Time

Abstract The study was conducted to detect Brucella sp. and Leptospira sp. in blood samples of dogs in Isfahan and Shahrekord province in Iran. A total of 94 blood samples were collected from dogs of different breed, age, sex, and dogs’ type (stray or nonstray). The samples were examined using conventional polymerase chain reaction (PCR). Fourteen (14.89%) dogs were positive for Brucella sp. and 18 (19.15%). dogs for Leptospira sp. There were no significant differences between the prevalence of the pathogens, provinces, sex, and age groups (P > 0.05). However, there was a statistically significant difference in prevalence of Brucella sp. and Leptospira sp. between stray and non-stray dogs (P < 0.0001; χ2 = 30.3767). The study also demonstrated that PCR was successfully used for the first time in Iran for the detection of Brucella sp. and Leptospira sp. in blood samples of dogs. Therefore, we recommend the PCR as a supplementary method with other commonly recognised methods (e.g. serological methods) for the diagnosis of subclinical infections with the microorganisms. Strict measures for the control of stray dogs are also highly recommended.

scholarly journals Molecular detection of lumpy skin disease virus in cattle by polymerase chain reaction in Iraq

2016 ◽  
Vol 40 (1) ◽  
pp. 83-88 ◽  
Author(s):  
Khitam Mahdi Mhemid
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Skin Disease ◽  
Age Groups ◽  
Conventional Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Lumpy Skin Disease Virus ◽  
Polymerase Chain

     This study is conducted to detect lumpy skin disease virus in Babylon, Al-Qadysia and Al-Muthana governorate during autumn 2014 using conventional polymerase chain reaction. A total of 150 specimens: 50 whole blood samples, 50 skin nodular biopsies and 50 tick samples were collected from infected animals of different breeds, genders and ages during lumpy skin disease outbreak. The results revealed that 104 cases (69.33%) were positive for lumpy skin disease virus by using polymerase chain reaction, with significant (P<0.05) differences between positive and negative cases. Out of 50 blood samples, 22 cases (44%) were positive for lumpy skin disease virus. The nodular skin samples collected from slaughtered animals showed 36 positive cases (72%), whereas 46 tick samples (92%) were positive for the disease, with significant (P<0.05) difference among them. According to gender, the finding showed significant results of lumpy skin disease in females (78.78%). It was recorded that higher percentage of positive cases was found in Friesian cattle (100%), crossbreed (73.58%) while native breed was (50.76%) with significant (P<0.05) difference among them. Regarding age groups, the results showed that all ages were susceptible to lumpy skin disease and significantly not different.

Molecular Detection of Leptospira Species Serotypes in Iranian Stray Dogs

2019 ◽  
Author(s):  
Saam Torkan ◽  
Hassan Momtaz
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Serum Samples ◽  
Blood Samples ◽  
Stray Dogs ◽  
Wide Range ◽  
Leptospira Spp ◽  
Polymerase Chain ◽  
Leptospira Species

Background and Aims: Leptospirosis is a spirochetal disease with public health importance globally. This disease affects a wide range of domestic and wild animals. Dogs are one of the species most sensitive to Leptospira canicola and Leptospira icterohaemorrhagiae. The present study was concluded to evaluate the prevalence rate of Leptospira species and L. canicola and L. icterohaemorrhagiae serovars in Iranian stray dogs. Materials and Methods: One-hundred and twenty blood samples were first taken from stray dogs. Then the samples were transferred to the laboratory. Sera were extracted from blood samples and genomic DNA was extracted. DNA samples were subjected to conventional polymerase chain reaction. Positive samples for Leptospira spp. were analyzed for presence of L. canicola and L. icterohaemorrhagiaeserovars using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: Nine samples out of 120 serum samples (7.5%) were positive for the flagella gene of the Leptospira spp. Prevalence of Leptospira spp. in serum samples of male and female dogs were 5.4% and 10.86%, respectively. Prevalence of L. canicola and L. icterohaemorrhagiae serovars were 55.55% and 33.33%, respectively. We found that 11.11% of samples were positive for both serovars. Two to three and 3-4 year old dogs had the highest prevalence of Leptospira spp. Conclusions: The considerable prevalence of leptospirta spp. and also their zoonotic serovars among Iranian stray dogs represented an important public health issue regarding the contact of healthy human with these dogs. Identification of infected dogs and their vaccination can inhibit the distribution of Leptospira spp.

scholarly journals Molecular detection of Babesia bovis in cattle in Al-Qadisiyah province

2017 ◽  
Vol 40 (2) ◽  
pp. 155-158
Author(s):  
Noaman N. A,aiz
Keyword(s):  
Polymerase Chain Reaction ◽  
Infection Rate ◽  
Molecular Detection ◽  
Babesia Bovis ◽  
Chain Reaction ◽  
Adult Cattle ◽  
Blood Samples ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Age And Sex

     This study aim to determine Babesia bovis infection in cattle based on genetic methods. A total of 96 blood samples were collected from alive and slaughtered cattle from different areas in addition to the abattoir of Al-Qadisiyah province from December 2013 to August 2014. Real time polymerase chain reaction (RT.PCR) technique was used to detect the presence of the protozoan with the effect of animal's age and sex in the infection rate 47.91 % (46/96) of examined cattle were given positive result to B. bovis infection. The highest infections were shown among the adult cattle (≥1 year), while there was non-significant difference (P>0.05) in the infection rate according to the sex. So the most cattle in Al-Qadisiyah province appear to be bearing the infection predominantly as a carrier hosts.

scholarly journals Polymorphisms in CCR5Δ32 and Risk of HIV-1 Infection in the Southeast of Caspian Sea, Iran

2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Zahra Heydarifard ◽  
Alijan Tabarraei ◽  
Abdolvahab Moradi
Keyword(s):  
Polymerase Chain Reaction ◽  
Hiv Infection ◽  
Caspian Sea ◽  
Chain Reaction ◽  
Blood Samples ◽  
Infected People ◽  
Significant Difference ◽  
Hardy Weinberg Equilibrium ◽  
Polymerase Chain ◽  
Hiv 1

Prevalence of CCR5Δ32 among blood samples of more than 400 healthy and HIV-1-infected people was investigated in Iran. Polymerase chain reaction (PCR) following DNA extraction was used. Desired frequency was analyzed by Hardy–Weinberg equilibrium (HWE) analysis and SPSS 16.0 software to harvest the results. The prevalence of CCRΔ32 heterozygote genotype was 3% in healthy people and 0.7% in HIV-1–infected individuals. There was no homozygote CCR5Δ32 in both groups, and the allele Δ32 was only observed in 1.5% and 0.36% of healthy and HIV-1–infected participants, respectively. Therefore according to this study, the frequency of the allele CCR5Δ32 indicates no significant difference between either groups (p=0.18) and it sounds that the mentioned mutation in heterozygote people would not affect their susceptibility against HIV infection. Genotyping trial in Iranians with HIV infection is supposed to be helpful as a matter of prognostic purposes.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Detection of MYCN Amplification in Serum DNA Using Conventional Polymerase Chain Reaction

2016 ◽  
Vol 31 (9) ◽  
pp. 1392 ◽  
Author(s):  
Youngeun Ma ◽  
Ji Won Lee ◽  
Soo Jin Park ◽  
Eun Sang Yi ◽  
Young Bae Choi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Conventional Polymerase Chain Reaction ◽  
Mycn Amplification ◽  
Chain Reaction ◽  
Serum Dna ◽  
Polymerase Chain

scholarly journals Critical analysis: use of polymerase chain reaction to diagnose leprosy

2016 ◽  
Vol 52 (1) ◽  
pp. 163-169 ◽  
Author(s):  
Flaviane Granero Maltempe ◽  
Vanessa Pietrowski Baldin ◽  
Mariana Aparecida Lopes ◽  
Vera Lúcia Dias Siqueira ◽  
Regiane Bertin de Lima Scodro ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Globin Gene ◽  
Public Health Problem ◽  
Reference Laboratory ◽  
Chain Reaction ◽  
Molecular Biology Technique ◽  
Pcr Inhibitor ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Pcr Techniques

ABSTRACT Leprosy is a neglected tropical disease and an important public health problem, especially in developing countries. It is a chronic infectious disease that is caused by Mycobacterium leprae, which has a predilection for the skin and peripheral nerves. Although it has low sensitivity, slit-skin smear (SSS) remains the conventional auxiliary laboratory technique for the clinical diagnosis of leprosy. Polymerase chain reaction (PCR) is a molecular biology technique that holds promise as a simple and sensitive diagnostic tool. In the present study, the performance of two PCR methods, using different targets, PCR-LP and PCR-P, were compared with SSS with regard to leprosy diagnosis in a reference laboratory. M. leprae DNA was extracted from 106 lymph samples of 40 patients who had clinical suspicion of leprosy. The samples were subjected to both PCR techniques and SSS. Amplification of the human b-globin gene was used as PCR inhibitor control. The specificity of both PCR techniques was 100%, and sensitivity was 0.007 and 0.015 µg/ml for PCR-LP and PCR-P, respectively. No significant difference was found between either the PCR-LP or PCR-P results and SSS results (p > 0.05). Although PCR is not yet a replacement for SSS in the diagnosis of leprosy, this technique may be used as an efficient auxiliary tool for early detection of the disease, especially in endemic regions. This strategy may also be useful in cases in which SSS results are negative (e.g., in paucibacillary patients) and cases in which skin biopsy cannot be performed.

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

Detection of a few DNA copies by real-time electrochemical polymerase chain reaction

The Analyst ◽  
2017 ◽  
Vol 142 (18) ◽  
pp. 3432-3440 ◽  
Author(s):  
M. Moreau ◽  
S. Delile ◽  
A. Sharma ◽  
C. Fave ◽  
A. Perrier ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Current Work ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
First Time ◽  
Accurate Quantification

In the current work, accurate quantification over 10 to 108 DNA copies has been successfully achieved for the first time by real-time electrochemical PCR.

scholarly journals Molecular identification by multiplex polymerase chain reaction of Pasteurella multocida in cattle and buffaloes in Baghdad

2014 ◽  
Vol 38 (1) ◽  
pp. 99-106
Author(s):  
Ihab G. M. AL-Shemmari
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Pasteurella Multocida ◽  
Type B ◽  
Chain Reaction ◽  
Blood Samples ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
B 31

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .

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