Identification of Pasteurella multocida by polymerase chain reaction

2020 ◽  
Vol 3 ◽  
pp. 14-18
Author(s):  
A.N. Seminа ◽  
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...  
Keyword(s):  
Pasteurella Multocida ◽  
Economic Losses ◽  
Sequence Database ◽  
Specific Determination ◽  
Polymerase Chain ◽  
Sick Bird ◽  
Positive Results ◽  
Highly Virulent ◽  
Poultry Farming

Pasteurellosis having a fairly wide distri-bution can be a reason that hinders the suc-cessful development of poultry farming. The causes and conditions for the occurrence of this disease in farms are often not specific. Pasteurella multocida can manifest itself from an extremely weakened Pasteurella carrier to a highly virulent pathogen. A sick bird is a hidden carrier of this disease and is subject to further culling. All this ultimately leads to significant economic losses. Rapid and reliable detection of this patho-gen will reduce or completely prevent eco-nomic losses. The clinical manifestation of the disease in the form of a lightning course causes difficulties in its lifetime diagnosis. The aim of our work was to develop unique samples of oligonucleotide sequences of primers specific to the pathogen P. multo-cida. The proposed method will provide de-tection of Pasteurella multocida within 3-4 hours, as well as rapid and highly specific determination of the type of this bacterium. After analyzing the Pasteurella multo-cida genome, we selected a region of the ptfA gene for the construction of oligonu-cleotides. A pair of primers were selected for PCR: Pm0567 and Pm1321. Gene se-quences were aligned by selecting similar ones in absolutely all the studied Pasteurel-la multocida isolates. Using special com-puter programs, the specificity of the select-ed primers was evaluated. A search in the sequence database revealed 100% homolo-gy of the selected primers with only homol-ogous sequences in the Pasteurella multo-cida genome. The study of samples contain-ing pathologic agents of bacterial nature by PCR confirmed the specificity of the select-ed primers. Positive results were obtained only with samples containing Pasteurella multocida. Thus, the selected primers can be proposed for the study of samples of different compositions to identify the ge-nome of Pasteurella multocida.

scholarly journals Methods of mycobacterial DNA isolation from different biological material: a review

2012 ◽  
Vol 51 (No. 5) ◽  
pp. 180-192 ◽  
Author(s):  
J. Hosek ◽  
P. Svastova ◽  
M. Moravkova ◽  
I. Pavlik ◽  
M. Bartos
Keyword(s):  
Dna Isolation ◽  
Economic Losses ◽  
Human Beings ◽  
Chain Reaction ◽  
Serious Infections ◽  
Biochemical Characterisation ◽  
Long Time ◽  
Polymerase Chain ◽  
Wide Family

Mycobacteria cause serious infections in animals and human beings. Huge economic losses on farms are caused by selected species of this wide family. A high risk of transmission of infection from animal to human exists. The knowledge of exact pathogen characteristics is an important factor which can improve quick and adequate healing. Cultivation and determination of phenotype is still the “gold standard”, but has the disadvantage of taking a long time and also low detection limit. Biochemical characterisation of isolates is not exact, and it is expensive. A more popular method used is the amplification of specific loci by polymerase chain reaction (PCR). For this method, the isolation of sufficient amounts of purified DNA is necessary. In this paper the most frequently used method for DNA isolation from live mycobacterial cells, body fluids, tissues, histological samples and forensic materials are outlined. This paper assists only as guide for these methods, so we describe them briefly.

Microbiological Monitoring of Sheep Carcass Contamination in Three Swiss Abattoirs

2003 ◽  
Vol 66 (6) ◽  
pp. 946-952 ◽  
Author(s):  
C. ZWEIFEL ◽  
R. STEPHAN
Keyword(s):  
Significant Relationship ◽  
Hazard Analysis ◽  
Sampling Site ◽  
Control Point ◽  
Critical Control Point ◽  
Plate Counts ◽  
Polymerase Chain ◽  
Carcass Contamination ◽  
Positive Results

At three Swiss abattoirs, 580 sheep carcasses were examined at 10 sites by the wet-dry double-swab technique. The aim of this study was to obtain data on microbiological contamination at the abattoirs and to develop a procedure for monitoring slaughter hygiene. Median aerobic plate counts (APCs) (log CFU/cm2) ranged from 2.5 to 3.8, with the brisket and neck sites showing the most extensive contamination. Enterobacteriaceae were detected on 68.1% of the carcasses and in 15.2% of the samples. The proportion of positive results ranged from 2.6% (for the hind leg and the flank at abattoir C) to 42.2% (for the perineal area at abattoir A). The percentage of samples testing positive for stx genes by polymerase chain reaction was 36.6%. A significant relationship between APC and the detection of Shiga toxin–producing Escherichia coli (STEC) was found for abattoirs A and B (depending on sampling site), whereas a significant relationship between Enterobacteriaceae and STEC detection was confirmed only for abattoir A (P < 0.05). In 57.1% of the 56 isolated non-O157 strains, stx2 genes were detected, and most of them were stx2d positive. Additional virulence factors were detected in 50% of the STEC strains, with 8.9% of these strains being eae positive, 50% being EHEC-hlyA positive, and 3.6% being astA positive. For the determination of carcass contamination, the monthly examination of 10 sheep carcasses for APC and Enterobacteriaceae counts in the neck, brisket, and perineal areas is recommended. This procedure is a valuable tool for the verification of slaughter hygiene according to hazard analysis critical control point principles.

Prevalence of SHV-extended spectrum β-lactamase producing carbapenem –resistantKlebsiellapneumoniae among patients with lower respiratory tractinfections in Babylon Province-Iraq.

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Resistance Rate ◽  
Combination Method ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Extended Spectrum ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections ◽  
Positive Results

This study was carried out to screen the prevalence of Klebsiella pneumoniae isolated from patients with lower respiratory tract infections in Babylon province.From December,2015 to the end of March,2016,a total of 100 sputum samples were collected from patients visited or hospitalized Merjan Teaching Hospital and Al- Hashimya General Hospital. Fifteenth (65%) isolates were identified as Klebsiellapneumoniae. All bacterial isolates were evaluated for extended spectrum β-lactamase (ESBL) production phenotypically using disk combination method. Eleven (73.3%) isolates were detected as ESBL-producers. Kirby-Bauer disk diffusion method was employed to determine resistance profile of ESBLs-positive isolates. Higher rates of resistance were observed for ampicillin and piperacillin antibiotics with (81.8%) and (72.7%) resistance rate, respectively, while the lowest rate was noticed for imipenem antibiotic (14.28%). Carbapenem-resistant isolates were investigated for blaSHV gene by Polymerase Chain Reaction (PCR) method, 2 (100%) isolates gave positive results.

Molecular Detection of Bacterial Pathogens Directly from the Nasal Swabs and Lung Tissue of Sheep and Goats

2019 ◽  
Vol 15 (02) ◽  
pp. 22-25
Author(s):  
Sunaina Thakur ◽  
Subhash Verma ◽  
Prasenjit Dhar ◽  
Mandeep Sharma
Keyword(s):  
Pasteurella Multocida ◽  
Direct Detection ◽  
Bacterial Species ◽  
Economic Losses ◽  
Isolation And Identification ◽  
Sheep And Goats ◽  
Histophilus Somni ◽  
Nasal Swabs ◽  
Mycoplasma Spp

Respiratory infections of sheep and goats cause heavy morbidity and mortality, leading to huge economic losses. Conventional methods of diagnosis that include isolation and identification of incriminating microbes are time-consuming and fraught with logistic challenges. Direct detection of incriminating microbes using molecular tools is gaining popularity in clinical, microbiological settings. In this study, a total of 50 samples (44 nasal swabs and 6 lung tissues) from sheep and goats were screened for the detection of different bacterial species by in vitro amplification of genus or species-specific genes. Histophilus somni was detected in 2% goat samples, Trueperella pyogenes in 20% goat nasal swabs, whereas 22% goat nasal swab samples were found positive for Mycoplasma spp. None of the samples from sheep was detected positive for H. somni, T. pyogenes, Mycoplasma spp. Similarly, all samples, irrespective, whether from sheep or goats, showed negative results for Pasteurella multocida, Mannheimia haemolytica, and Corynebacterium pseudotuberculosis.

scholarly journals Complicated cutaneous leishmaniasis caused by an imported case of Leishmania tropica in Japan: a case report

2021 ◽  
Vol 49 (1) ◽  
Author(s):  
Hiroyuki Kitano ◽  
Chizu Sanjoba ◽  
Yasuyuki Goto ◽  
Kazumasa Iwamoto ◽  
Hiroki Kitagawa ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Skin Lesions ◽  
Leishmania Tropica ◽  
Case Presentation ◽  
Imported Case ◽  
Imported Cases ◽  
History Of ◽  
Hematoxylin And Eosin ◽  
Polymerase Chain ◽  
Positive Results

Abstract Background Leishmaniasis is not endemic in Japan, and imported cases are rare. However, there are increasing concerns regarding imported cases of cutaneous leishmaniasis from endemic countries to Japan. This report describes a case of imported cutaneous leishmaniasis that was diagnosed and treated in Japan. Case presentation A 53-year-old Pakistani man presented with skin lesions on both malleoli of his right ankle and the dorsum of the left foot. The skin lesions manifested as erythematous nodules surrounding an ulcer in the center of the lesion. The lesions of the malleoli of his right ankle each measured 3 × 3 cm, and the lesion on the top of his left foot measured 5 × 4 cm. He had been living and working in Japan but had a history of a visit to Pakistan for about 2 months in 2018. The skin lesions were biopsied. Giemsa and hematoxylin and eosin staining of biopsy samples showed amastigotes of Leishmania in macrophages, and the presence of Leishmania was confirmed by skin tissue culture. Polymerase chain reaction using biopsy specimens identified Leishmania parasites, and DNA sequence analysis revealed that the species was Leishmania tropica. The patient was treated with intravenous liposomal amphotericin B for 6 days. The erythema disappeared, and the erythematous nodules resolved within 3 weeks. Conclusion This is the first report of imported cutaneous leishmaniasis caused by L. tropica from Pakistan, and it is interesting that all three testing modalities showed positive results in this case.

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