Identification of Leptospira in Patients With Renal Failure Using Serological, Molecular and Pathological Based Techniques, in Shiraz, Iran

Abstract Leptospirosis is a relatively rare bacterial infection that affects people and animals caused by pathogenic species of Leptospira. The present study was conducted using nested polymerase chain reaction (PCR), microscopic agglutination test (MAT) and hematological and biochemical tests on 200 blood samples of renal disorders patients in Shiraz, Iran. Also nested-PCR assay and Warthin-Starry (WS) silver staining method was performed on 30 nephrectomised kidney sample. The frequency of pathogenic species of Leptospira infection in patients with renal disorders was 20 % and this infection was significantly correlated with BUN, anemia, RDW, MCV, MCH and hemoglobin levels (P < 0.01). MAT analysis showed that serum samples had positive titers against L. Grippotyphosa (13 samples), L. Ballum (6 sample), L. Pomona (3 samples), L. Canicola (2 samples), L. Icterohaemorrhagiae (1 sample) and L. Hardjo (1 sample) serovars. Twenty-three percent of the kidney samples from the patients with pyelonephritis were infected with the pathogenic species of Leptospira. This study showed that pathogenic Leptospira serovars are present in this area and in patients with renal disorders more attention should be paid to this zoonotic disease.

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Parvovirus B19 infection in anemic children with AIDS using a nested polymerase chain reaction (PCR) assay: correlation with slot blot hybridization and antibody status by enzyme linked immunosorbent assay (ELISA)† 911

Pediatric Research ◽

10.1203/00006450-199604001-00933 ◽

1996 ◽

Vol 39 ◽

pp. 154-154

Author(s):

Ninad Desai ◽

Elizabeth Secord ◽

Marc S.C Cheah ◽

Sreedhar P Rao

Keyword(s):

Parvovirus B19 ◽

Blot Hybridization ◽

Enzyme Linked Immunosorbent Assay ◽

Slot Blot ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Slot Blot Hybridization ◽

Parvovirus B19 Infection ◽

Polymerase Chain

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Ocorrência de anticorpos contra Neospora caninum em bovinos e cães oriundo de propriedades produtoras de leite do Norte Central do Estado do Paraná

Semina Ciências Agrárias ◽

10.5433/1679-0359.2018v39n6p2449 ◽

2018 ◽

Vol 39 (6) ◽

pp. 2449

Author(s):

Simone Marins Spiti ◽

Luiz Daniel de Barros ◽

Mércia De Seixas ◽

Ana Flavia Minutti ◽

Thais Agostinho Martins ◽

...

Keyword(s):

Neospora Caninum ◽

Rare Event ◽

Epidemiological Studies ◽

Northern Region ◽

Clot Retraction ◽

Chain Reaction ◽

Serum Samples ◽

Nested Polymerase Chain Reaction ◽

Blood Samples ◽

Polymerase Chain

Neosporosis is one of the principal causes of reproductive problems in cattle worldwide. The aim of this study was to evaluate the occurrence of anti-Neospora caninum antibodies in cattle and dogs from dairy farms from the central northern region of Paraná state. Blood samples with and without EDTA were collected from 400 cattle and 46 dogs from 20 properties. Nested polymerase chain reaction (n-PCR) was performed on whole blood samples and indirect immunofluorescence reaction (IFR) on the serum samples (after clot retraction). Cattle and dogs with titers ? 100 and ? 50, respectively, were considered positive. Anti-N. caninum was detected in 20,1% (80/400) of cattle, with titers ranging from 100 to 1600. The n-PCR presented only two cattle positives (0.5%). Anti-N. caninum was detected in 19,6% (9/46) of the dogs, with titers ranging from 50 to 6400. The occurrence of antibodies against N. caninum obtained in the present study was similar to those in studies performed in other regions of Paraná and Brazil. The probability of detecting parasitemia in epidemiological studies is a rare event.

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Sensitivity of the nested-polymerase chain reaction (PCR) assay for Brugia malayi and significance of ‘free’ DNA in PCR-based assays

International Journal for Parasitology ◽

10.1016/s0020-7519(00)00104-1 ◽

2000 ◽

Vol 30 (11) ◽

pp. 1177-1179 ◽

Cited By ~ 6

Author(s):

Janet Cox-Singh ◽

Andrea S Pomrehn ◽

Nathan D Wolfe ◽

Hasan A Rahman ◽

Huong-Ying Lu ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Brugia Malayi ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Free Dna ◽

Polymerase Chain

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Digenetic trematodes,Acanthatriumsp. andLecithodendriumsp., as vectors ofNeorickettsia risticii, the agent of Potomac horse fever

Journal of Helminthology ◽

10.1079/joh2003181 ◽

2003 ◽

Vol 77 (4) ◽

pp. 335-339 ◽

Cited By ~ 38

Author(s):

N. Pusterla ◽

E.M. Johnson ◽

J.S. Chae ◽

J.E. Madigan

Keyword(s):

Aquatic Insects ◽

Preliminary Evidence ◽

Tachycineta Bicolor ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Digenetic Trematodes ◽

Potomac Horse Fever ◽

The Family ◽

Polymerase Chain

AbstractNeorickettsia(formerlyEhrlichia)risticii, the agent of Potomac horse fever (PHF), has been recently detected in trematode stages found in the secretions of freshwater snails and in aquatic insects. Insectivores, such as bats and birds, may serve as the definitive host of the trematode vector. To determine the definitive helminth vector, five bats (Myotis yumanensis) and three swallows (Hirundo rustica,Tachycineta bicolor) were collected from a PHF endemic location in northern California. Bats and swallows were dissected and their major organs examined for trematodes and forN. risticiiDNA using a nested polymerase chain reaction (PCR) assay. Adult digenetic trematodes,Acanthatriumsp. and/orLecithodendriumsp., were recovered from the gastrointestinal tract of all bats and from one swallow. The intestine of three bats, the spleen of two bats and one swallow as well as the liver of one swallow tested PCR positive forN. risticii. From a total of seven pools of identical digenetic trematodes collected from single hosts, two pools ofAcanthatriumsp. and one pool ofLecithodendriumsp. tested PCR positive. The results of this investigation provide preliminary evidence that at least two trematodes in the family Lecithodendriidae are vectors ofN. risticii. The data also suggest that bats and swallows not only act as a host for trematodes but also as a possible natural reservoir forN. risticii.

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Evaluation and Use of a Nested Polymerase Chain Reaction Assay in Cats Experimentally Infected with Bartonella Henselae Genotype I and Bartonella Henselae Genotype II

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870101300406 ◽

2001 ◽

Vol 13 (4) ◽

pp. 312-322 ◽

Cited By ~ 7

Author(s):

Alma F. Roy ◽

Richard E. Corstvet ◽

Ronald A. Tapp ◽

Kathy L. O'Reilly ◽

Hollis U. Cox

Keyword(s):

Polymerase Chain Reaction ◽

Bartonella Henselae ◽

Turnaround Time ◽

Nucleic Acid Amplification ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Genotype I ◽

Polymerase Chain ◽

Genotype Ii

Cats have been shown to be infected with Bartonella henselae genotype I, B. henselae genotype II, and B. clarridgeiae. Feline bartonellosis infections and the strains involved in these infections are important in both veterinary and human medicine. Nucleic acid amplification methods such as polymerase chain reaction (PCR) are being used in both research and diagnostics as tools for understanding many infectious diseases. Bartonella bacteremia in cats is detected by blood culture; however, because of the limitations of culture (delayed turnaround time and sensitivity limits), PCR may be a more efficient method for identifying infected cats. Three distinct PCR assays that could differentiate among B. henselae genotype I, B. henselae genotype II, and B. clarridgeiae were developed and used to detect as few as 3.2 organisms. Fourteen cats experimentally infected with B. henselae genotype I and B. henselae genotype II were followed by bacterial culture and PCR through the course of infection, including periods of primary and relapsing bacteremia. The PCR assay was positive in 11 of the 14 cats for periods of 1–9 weeks after culture became negative. Of the 223 blood specimens that were culture negative, the PCR assay was positive in 38 (17%) of the specimens. Two of the 14 cats developed relapsing bacteremia. The 2 B. henselae genotypes were amplified in the cats and the bacteremic phase of these infections as determined by PCR lasted for a longer period than previously determined by culture. Using laboratory assays such as PCR to understand the strains involved in feline bartonellosis and the course of the infection is important in the understanding of these zoonotic agents.

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Assessment of the First Commercial ELISA Kit for the Diagnosis ofTheileria annulata

Journal of Parasitology Research ◽

10.1155/2015/787812 ◽

2015 ◽

Vol 2015 ◽

pp. 1-4 ◽

Cited By ~ 1

Author(s):

Amira A. T. Al-Hosary ◽

Jabbar Ahmed ◽

Ann Nordengrahn ◽

Malik Merza

Keyword(s):

Surface Protein ◽

Theileria Annulata ◽

Reference Test ◽

Chain Reaction ◽

Pcr Assay ◽

Serum Samples ◽

Elisa Kit ◽

Epidemiological Surveys ◽

Merozoite Surface Antigen ◽

Polymerase Chain

The present study assesses the efficacy of SVANOVIRTheileria annulata-Ab, the first commercial ELISA kit for the diagnosis ofTheileria annulatainfection in cattle based on a recombinant protein known asT. annulatasurface protein (TaSp). As a reference test, a polymerase chain reaction (PCR) assay depending onT. annulatamerozoite surface antigen (Tams-1) was applied. A total of 468 blood samples as well as serum samples were randomly collected from cattle and tested in the PCR as well as in the ELISA developed in this study. Moreover, all samples were also analyzed by conventional Giemsa-stained blood smear. The results of this study revealed a good correlation between the results obtained by PCR and the ELISA, whereas all PCR positive samples scored correctly positive in the ELISA and 73 of the 125 PCR negative samples scored correctly negative. Taken together, a sensitivity of 91.25% and a specificity of 78.4% were recorded, when compared to the PCR data. In conclusion, the SVANOVIRTheileria annulata-Ab is a suitable diagnostic assay for use in the diagnosis and epidemiological surveys ofTheileria annulatainfection in chronic and carrier animals.

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A Novel Nested Polymerase Chain Reaction (n-PCR) Assay for Identifying Sorghum nitidum

Notulae Scientia Biologicae ◽

10.15835/nsb346329 ◽

2011 ◽

Vol 3 (4) ◽

pp. 143-146

Author(s):

Shasha WEI ◽

Zhirui DENG ◽

Liping YIN ◽

Jianping YI ◽

Renqi WU ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Nucleotide Sequence ◽

Sorghum Bicolor ◽

Plant Protection ◽

Sorghum Halepense ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Field Inspection ◽

Polymerase Chain

This work developed a novel nested polymerase chain reaction (n-PCR) assay to identify Sorghum nitidum (S. nitidum). It has been designed a set of specific n-PCR inner primers Snit5/Snit2 and outer primers Nout1/Nout2 based on a conserved nucleotide sequence of adh1-like gene of S. nitidum. Fourteen samples of sorghum were used to investigate the specificity of the primers and the n-PCR assay. The result showed that 9 samples of S. nitidum displayed a positive strong, specific amplified band at ~873 bp in gel spectra, while other relatives, including Sorghum halepense, Sorghum almum, Sorghum bicolor, Sorghum propinum and Sorghum sudanse exhibited negative amplifications. This assay was able to specifically identify S. nitidum fast and effectively, which could be applied widely in field inspection, agriculture production and plant protection.

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Development of a Nested Polymerase Chain Reaction Assay for Detection of Colletotrichum acutatum on Symptomless Strawberry Leaves

Plant Disease ◽

10.1094/pdis-92-12-1655 ◽

2008 ◽

Vol 92 (12) ◽

pp. 1655-1661 ◽

Cited By ~ 9

Author(s):

O. Pérez-Hernández ◽

M. H. Nam ◽

M. L. Gleason ◽

H. G. Kim

Keyword(s):

Polymerase Chain Reaction ◽

Colletotrichum Acutatum ◽

Tween 20 ◽

Chain Reaction ◽

Pcr Assay ◽

Nested Polymerase Chain Reaction ◽

Plastic Bags ◽

Reliable Diagnosis ◽

Polymerase Chain ◽

Composite Samples

A nested polymerase chain reaction (PCR) assay was developed for detection of Colletotrichum acutatum on symptomless strawberry leaves. In pure culture, the assay detected as little as 1.0 fg of DNA extracted from mycelium and as few as 1.5 conidia ml–1 when conidial suspensions were sonicated. On detached inoculated leaves, three alternative protocols to dislodge the pathogen were assessed: (i) immersion of whole leaves in 0.05% Tween 20 and manual agitation in plastic bags for 1 min (A); (ii) immersion in Tween 20, sonication for 30 min, then agitation for 1 min (SA); and (iii) freezing for 3 h, incubation for 2 days at 27°C, immersion in Tween 20, then sonication for 30 min and agitation for 1 min (FISA). Each method removed significantly (P ≤ 0.05) more conidia from leaves than the nontreated control; however, removal of appressoria did not vary among assays. In composite samples of noninoculated and inoculated (1.5 ×103 conidia ml–1) strawberry leaves, the nested PCR assay using the FISA protocol detected C. acutatum in as few as 1 infested leaf in 50 noninfested leaves. In a strawberry field, the assay detected the presence of C. acutatum in samples of asymptomatic strawberry leaves, showing potential as a powerful tool for reliable diagnosis of the pathogen in the field.

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