Development of a novel molecular assay based on conventional PCR liquid hybridization and ELISA (NAT-ELISA) for the detection of HIV, HCV, and HBV in blood donors

Ascaridiosis is a very important parasitic disease of birds, it is caused by Ascaridia. This study was conducted to identify the Ascaridia species by microscopic and molecular assay in Baghdad city. One hundred and sixty fecal samples were collected from domestic pigeons during the period from 1/1/ 2019 to 31/3/ 2019. Results showed that the rate of infection for Ascaridia spp. 15.62% by microscopic examination. Significant difference was observed in infection rates between males and females pigeons. Fifty samples randomly selected and subjected to molecular diagnosis of Ascaridia spp.. Molecular examination results, the total infection rate showed 16%(8/50). The eight positive PCR products were sequenced and deposited in Gene bank data base, phylogenic analysis demonstrated that 4 sequences belongs to Ascaridia galli ( MK918635.1, MK918636.1, MK918847.1, MK919081.1), while 2 (MK919199.1, MK919200.1) belong to Ascaridia nymphii and 2 (MK919207.1, MK919264.1) belong to Ascaridia numidae. It is the first study in Iraq to diagnosis of Ascaridia nymphii and Ascaridia numidae in domesticed pigeons by using conventional PCR.

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Prevalence of Haemoplasma Infections in Stray Cats in Northern Italy

ISRN Microbiology ◽

10.1155/2014/298352 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 14

Author(s):

Eva Spada ◽

Daniela Proverbio ◽

Paola Galluzzo ◽

Alessandra Della Pepa ◽

Giada Bagnagatti De Giorgi ◽

...

Keyword(s):

Risk Factors ◽

Blood Donors ◽

Northern Italy ◽

Negative Association ◽

Domestic Cats ◽

Conventional Pcr ◽

Blood Samples ◽

Stray Cats ◽

Stray Cat

This study investigated the prevalence of feline haemoplasma infections in a number of stray cat colonies in Milan, Northern Italy. Blood samples from 260 stray cats were evaluated, with conventional PCR, for the presence of DNA associated with Mycoplasma haemofelis (Mhf) and “Candidatus Mycoplasma haemominutum” (CMhm). Odd ratios (OR) were calculated to identify risk factors for haemoplasma infections. PCR was positive in 86 out of 260 subjects (33.1%), with a prevalence of 10.8% (28/260 cats) for Mhf and 22.3% (58/260 cats) for CMhm. No coinfections were registered. There were significant associations between infections and season of sampling, that is, a negative association between winter sampling and a haemoplasma positive status (OR=0.29, P=0.001), or CMhm positive status (OR=0.29, P=0.01). Haemoplasma infections are common in stray cats in Milan. Thus, domestic cats with outdoor access should be routinely monitored and treated for ectoparasites to minimize risks of disease acquisition. Moreover, as these infections are transmitted via blood, feline blood donors from this area should be screened by PCR and preferably be drawn from a population of indoor cats regularly treated for fleas.

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Molecular Assay Proves the Presence of Theileria annulata Infection in Camels in Al-Diwaniyah Province, Iraq

Iranian Journal of Parasitology ◽

10.18502/ijpa.v16i2.6317 ◽

2021 ◽

Author(s):

Noaman N. A'aiz ◽

Hayder N. Ayyez ◽

Ahmed J. Neamah

Keyword(s):

Dna Amplification ◽

18S Rrna Gene ◽

Rrna Gene ◽

Molecular Assay ◽

Conventional Pcr ◽

Molecular Investigation ◽

Causative Agents ◽

The One ◽

Specific Species ◽

Theileria Spp

Background: Theileria camelensis and T. dromedarii are parasitic protozoans reported by several studies as specific species that infect the one-humped camel (Camelus dromedarius). However, other findings casted significant doubts on the true identity of the causative species of theileriosis in camels. Therefore, the present study was conducted to investigate of T. camelensis and T. dromedarii in one humped camels in Iraq during Apr-Oct 2017. Methods: Blood samples for DNA extraction were obtained from 181 slaughtered camels. Molecular investigation was performed following the amplification of 18S rRNA gene by conventional PCR technique. DNA sequencing was then utilized only for the positive samples to confirm the infection with the Theileria species. Results: Nine (4.97%) out of 181 examined samples showed a positive result to infection with Theileria spp., and all these appeared as a T. annulata when subjected to DNA amplification and sequencing techniques. There was a complete absence of any new sequence outside the known species. Conclusion: Most of Theileria infection in camels in the study area is caused by T. annulata and no other causative agents like T. camelensis or T. dromedarii.

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Choice of molecular assay determines ranavirus detection probability and inferences about prevalence and occurrence

Diseases of Aquatic Organisms ◽

10.3354/dao03518 ◽

2020 ◽

Vol 141 ◽

pp. 139-147

Author(s):

FJ Wynne ◽

R Puschendorf ◽

ME Knight ◽

SJ Price

Keyword(s):

Population Declines ◽

Molecular Assay ◽

Conventional Pcr ◽

Emerging Pathogens ◽

Pcr Assay ◽

Molecular Assays ◽

Field Samples ◽

Assay Precision ◽

Viral Lineage ◽

Inter Assay Variation

Ranaviruses are emerging pathogens that can cause morbidity, mortality and population declines in ectothermic hosts; however, there is no standardized approach to diagnostics. Here, we compared the inter-assay variation and intra-assay precision among 2 commonly used quantitative PCRs (qPCRs), a conventional and a nested PCR assay (used as a gold standard), using laboratory-propagated ranavirus (FV3 and CMTV) and field-collected samples. A qPCR assay (‘Leung’) detected viral DNA in dilutions 2 orders of magnitude lower than other assays regardless of the viral lineage of the cultured isolate (FV3/CMTV). The second qPCR (‘Brunner’) was slightly more sensitive than the conventional PCR (‘Mao’ assay). For field samples, the Leung qPCR detected all known positives, while the Mao assay PCR only detected 2.5% of the positive samples. Amplicon sequences from the 2 conventional PCRs were shown to be useful for inferring viral lineage. Inaccurate results will bias estimates of the distribution and prevalence of ranaviruses, and together these findings emphasize that molecular assays should be chosen carefully in the context of study aims.

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Identification of Leishmania infantum in blood donors from endemic regions for visceral leishmaniasis

Parasitology ◽

10.1017/s0031182020001936 ◽

2020 ◽

pp. 1-5

Author(s):

Loren Queli Pereira ◽

Márcia Maria Ferreira-Silva ◽

Cristhianne Molinero Andrade Ratkevicius ◽

César Gómez-Hérnandez ◽

Fernanda Bernadelli De Vito ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Leishmania Infantum ◽

Blood Donors ◽

Asymptomatic Infection ◽

Molecular Techniques ◽

Enzyme Linked Immunosorbent Assay ◽

Conventional Pcr ◽

Asymptomatic Patients ◽

Polymerase Chain ◽

Asymptomatic Individuals

Abstract Visceral leishmaniasis is an endemic protozoonosis observed in over 60 countries, with over 500 000 new cases recorded annually. Although the diagnostic procedure of its symptomatic forms is well established, for asymptomatic patients, who represent about 85% of those infected, there is no consensus on the best method for its identification. Recent studies have presented molecular techniques as viable identification methods, with good sensitivity and specificity indices in asymptomatic individuals. Therefore, we aimed to use molecular methods to assess their effectiveness in identifying the presence of asymptomatic infection by Leishmania infantum (L. infantum) individuals from endemic regions of Brazil. Screening was performed by real-time polymerase chain reaction (qPCR) and confirmed by sequencing the cytochrome B gene. Of the 127 samples [from 608 blood donors who had participated in a previous study, of which 34 were positive by the enzyme-linked immunosorbent assay (ELISA) rK39] tested by qPCR, 31 (24.4%) were positive. In the sequencing of 10 qPCR-positive samples, five were identified as L. infantum. Complimentary samples of the ELISA rK39 and conventional PCR showed only reasonable and low agreement with qPCR, respectively. The qPCR confirmed the presence of infection in five of the 10 sequenced samples, ELISA confirmed three, and the conventional PCR confirmed none.

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Hemoplasma prevalence and hematological abnormalities associated with infection in three different cat populations from Southern Brazil

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612014079 ◽

2014 ◽

Vol 23 (4) ◽

pp. 428-434 ◽

Cited By ~ 13

Author(s):

Andrea Pires dos Santos ◽

Francisco de Oliveira Conrado ◽

Joanne Belle Messick ◽

Alexander Welker Biondo ◽

Simone Tostes de Oliveira ◽

...

Keyword(s):

Blood Donors ◽

Blood Collection ◽

Southern Brazil ◽

Domestic Cats ◽

Conventional Pcr ◽

Different Populations ◽

Hematological Abnormalities ◽

Complete Blood Counts ◽

Candidatus Mycoplasma Turicensis ◽

Hematological Alterations

Three hemoplasma species are recognized in domestic cats: Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’ and ‘Candidatus Mycoplasma turicensis’. We report the prevalence and hematological abnormalities of hemoplasma infection in 369 domestic cats from three different populations (blood donors, hospitalized cats and shelter cats) from Southern Brazil. Complete blood counts were performed at the time of blood collection, and DNA was extracted and tested by conventional PCR for each hemoplasma species. A total of 79 samples (21.40%) were positive for at least one species. The most prevalent hemoplasma was ‘Candidatus Mycoplasma haemominutum’, with 50/369 (13.55%) positive cats, followed by ‘Candidatus Mycoplasma turicensis’, 10/369 (2.71%), and Mycoplasma haemofelis, 8/369 (2.16%). Mycoplasma haemofelis and ‘Candidatus Mycoplasma haemominutum’ coinfection was observed in 4/369 (1.08%), whereas ‘Candidatus Mycoplasma haemominutum’ and ‘Candidatus Mycoplasma turicensis’ in 5/369 (1.35%). Three cats (0.81%) were infected with all three hemoplasmas. There was no association between infection and the different populations. Anemia was associated with Mycoplasma haemofelis and ‘Candidatus Mycoplasma haemominutum’, but not with ‘Candidatus Mycoplasma turicensis’. Male cats and cats with outdoor access were more likely to be infected. Although ‘Candidatus Mycoplasma haemominutum’ is believed to cause minimal or no hematological alterations, the infected cats studied herein were more likely to be anemic.

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