scholarly journals Differential diagnosis of ovine endogenous and pulmonary oncogenic jaagsiekte retroviruses in Mongolia

2018 ◽  
Vol 25 (03) ◽  
pp. 49-56
Author(s):  
Undarmaa S ◽  
Davaasuren N ◽  
Nyamgarav B ◽  
Tserennadmid L ◽  
Batnasan T ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Long Terminal Repeat ◽  
Restriction Endonuclease ◽  
Lung Tumor ◽  
Terminal Repeat ◽  
Blood Samples ◽  
Specific Pcr ◽  
Pcr Product ◽  
Central Regions ◽  
Pcr Products

The endogenous jaagsiekte retrovirus (exoJSRV) is integrated into the sheep genome as result of long coevolution of the virus and host and transmitted vertically. The exogenous jaagsiekte retrovirus (endoJSRV) is transmitted from infected sheep and induce ovine pulmonary adenocarcinoma. The differential diagnosis of endoJSRV from the exogenous oncovirus (exoJSRV) is based on unique nucleotide sequences of exoJSRV which is characterized by a digestion of Sca I restriction endonuclease while endo JSRV not. We used the routine diagnostic method for determination of both JSRV in healthy and affected sheep in Mongolia as rest of the world. The 229 bp long PCR product of the Gag region of the virus was amplified in all 81 sheep blood samples from central regions but not digested by Sca I restriction endonuclease which indicate the presence of endoJSRV. The nucleotide sequence of two PCR products of khalkh sheep breed were clustered with viral strains from India and China, respectively. The lung tumor nodule was present in sheep with breeding problems, the hematoxylin and eosin staining of the affected lungs showed strong tumor cell proliferation sites and decreased number of alveoli. The 229 bp PCR product of viral Gag region was digested by Sca I restriction endonuclease which indicate the presence of exogenic JSRV, the oncovirus. The LTR (long terminal repeat) specific PCR of the exoJSRV resulted as positive in two tumor lung samples except non-tumor lung samples and blood samples which validated our results strongly. Монгол хонины эндоген болон уушигны хорт хавдар үүсгэгч онковирусийг ялган оношлосон нь Хураангуй: Хонины эндоген жаагсиектэ ретровирус (Jaagsiekte retrovirus -хJSRV/ЖРВ) нь олон жилийн турш эзэн амьтан болон вирусийн хам эволюцийн явцад эзэн амьтны геномд нэгдэн хонинд удамшдаг болсон байна. Харин ЖРВ-ийн эксоген хэлбэр нь өвчтэй малаас удамшлын бус замаар малд халдварлан уушигны хорт хавдар-аденоматозыг үүсгэдэг онковирус тул ЖРВ-ийн эдгээр хэлбэрийг ялган оношлох нь Монгол орны малын халдварт өвчний үнэн бодит шинжилгээнд зайлшгүй чухал юм. xЖРВ-ийн энэхүү 2 хэлбэр нь хоорондоо төстэй боловч нуклеотидийн дараалал, ялангуяа вирусийн gag хэсгийн 229 хос суурь урттай хэсэг нь Scal эндонуклеаз ферментээр таслагдах эсэхээрээ ялгагддаг нь бусад орны судлаачдын үр дүнгээр тогтоогдсон байна. Бидний цуглуулсан 81 толгой хонины цусны бүх дээжинд 229 хос суурь урттай ДНХ-ийн судал илэрсэн боловч ScaI эндонуклеаз ферментээр хэсэгчлэн таслагдаагүй мөн нуклеоитидийн дарааллыг секвенсингээр тогтооход энэ нь ЖРВ-ийн эндоген хувилбар болох нь нотлогдов. Эмнэлзүйн үзлэгээр амьсгаа нь давхацсан, уушиг хэржигнэсэн шуугиантай хонины уушигны баруун хэлтэр нь 2-3 дахин томорсон, цайвар өнгөтэй болж, хатуурсан, эмгэг бие бүтцийн шинжилгээгээр уушигны цулцангийн ханын шүүрлийн хучуур эс хорт хавдрын эс болон хувирч олшрон үржлийн голомтууд үүсгэн агаарын хийн солилцооны гуурс руу түрэн орж хөхөлгөр ур үүсгэсэн нь ажиглагдав. Энэ судалгаагаар бид эндонуклеаз фермент ашиглан уушигны аденоматоз, хорт хавдар үүсгэгч эксоген ЖРВ-ийн халдварыг бүх хонинд удамшдаг эндоген ЖРВ-ээс ялган оношлох боломжийг нотолсон арга боловсруулав. Мөн LTR хэсгийн өвөрмөц хос праймер ашиглан эксоген ЖРВ-йиг ПГУ-аар илрүүлэх боломжтойг баталгаажуулав. Түлхүүр үг: Жаагсиектэ ретровирус (ЖРВ), онковирус, эксоген ЖРВ, эндоген ЖРВ, хонины уушигны аденоматоз, эндонуклеаз фермент, gag, LTR-long terminal repeat, хөхөлгөр ур

Multiplex PCR for differential diagnosis of emerging typhoidal pathogens directly from blood samples

2008 ◽  
Vol 137 (1) ◽  
pp. 102-107 ◽  
Author(s):  
A. ALI ◽  
ABDUL HAQUE ◽  
ASMA HAQUE ◽  
Y. SARWAR ◽  
M. MOHSIN ◽  
...  
Keyword(s):  
Developing Countries ◽  
Differential Diagnosis ◽  
Blood Culture ◽  
Multiplex Pcr ◽  
Mixed Infection ◽  
Major Health Problem ◽  
Major Health ◽  
Blood Samples ◽  
Specific Pcr ◽  
Pcr Targeting

SUMMARYClassicallySalmonella entericaserovar Typhi (S. Typhi) is associated with typhoid, a major health problem in developing countries. However, in recent yearsS. Paratyphi A and Vi-negative variants ofS. Typhi have emerged rapidly. We have developed a nested multiplex PCR targeting five different genes for differential diagnosis of typhoidal pathogens which has been optimized to be directly applicable on clinical blood samples. Of 42 multiplex PCR-positive blood samples, 26, nine, and two were Vi-positiveS. Typhi, Vi-negativeS. Typhi andS. Paratyphi A, respectively, and five patients were found to have mixed infection. Seventeen patients grewSalmonellafrom blood culture and the remaining 25 were positive in theSalmonella-specific PCR. Tests with several common pathogens confirmed the specificity of the assay. We conclude that the proposed multiplex PCR is rapid, sensitive and specific for the diagnosis of typhoidal pathogens directly from blood samples.

Analysis of the inhibition of nucleic acid dyes on polymerase chain reaction by capillary electrophoresis

2016 ◽  
Vol 8 (11) ◽  
pp. 2330-2334 ◽  
Author(s):  
Zhenqing Li ◽  
Chenchen Liu ◽  
Siyao Ma ◽  
Dawei Zhang ◽  
Yoshinori Yamaguchi
Keyword(s):  
Polymerase Chain Reaction ◽  
Capillary Electrophoresis ◽  
Nucleic Acid ◽  
Acid Dyes ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Pcr Product ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Accurate Quantification

An integrated polymerase chain reaction (PCR) and capillary electrophoresis (CE) system can realize accurate quantification of the target PCR product by adding labeling dyes to the PCR reagents, because CE can discriminate all the subsequent nucleic acids, including the primers, non-specific and specific PCR products.

scholarly journals Distribution of the Beta-Casein Gene Variants in Three Cattle Breeds Reared in Benin

2022 ◽  
Vol 14 (2) ◽  
pp. 86
Author(s):  
Camus Mahougnon Adoligbe ◽  
Stéphanie Gloria Akpo ◽  
Santoze Adido ◽  
Marguéritte M’Po ◽  
Ange-Régis Zoclanclounon ◽  
...  
Keyword(s):  
The Other ◽  
Gene Variants ◽  
Blood Samples ◽  
Casein Gene ◽  
Average Percentage ◽  
Cattle Breeds ◽  
Specific Pcr ◽  
Pcr Products ◽  
Beta Casein ◽  
Good Potential

The beta-casein gene is one of the most functional genetic candidate that affect milk quality and composition traits. Among its variants, the A1/A2 are the most common. Therefore, the aim of this study was to identify the distribution of the Beta-casein gene variants (A1/A2) in three different cattle breeds in order to determine which of the breed produce a better milk for consumers’ health. 152 blood samples which comprises 72 (Muturu), 40 (Azawak) and 40 Girolando were used to carry out this study. Genomic DNA was extracted from the blood samples and each variant was subsequently amplified from the extracted DNA samples using an Allele-Specific PCR technique and then confirmed by running the PCR products on 1% agarose gel. The result showed that there were three genotypes (A1A1, A2A1 and A2A2) in the three breeds. The average percentage genotypic frequencies obtained from this study were 42.76%, 31.58% and 25.66% respectively for A1A1, A1A2 and A2A2 genotypes while the percentage allelic frequencies were 58% and 42% respectively for A1 and A2 allele. The genetic parameters of Azawak breed were higher than that of the other breeds, what implies that there was a higher polymorphism and genetic diversity in the Azawak breed in the beta-casein gene compare to the other breeds. The A2 beta-casein variant in milk has been found to be desirable for milk consumer’s health and nutrition. This study therefore showed that the Azawak breed provides a good potential for increasing this favorable allele through appropriate breeding techniques of cattle.

scholarly journals The long terminal repeat is a determinant of cell tropism of maedi-visna virus

2000 ◽  
Vol 81 (8) ◽  
pp. 1901-1905 ◽  
Author(s):  
Gudrún Agnarsdóttir ◽  
Holmfrídur Thorsteinsdóttir ◽  
Thórdur óskarsson ◽  
Sigrídur Matthíasdóttir ◽  
Benedikta St. Haflidadóttir ◽  
...  
Keyword(s):  
Long Terminal Repeat ◽  
Pcr Amplification ◽  
Cell Types ◽  
Terminal Repeat ◽  
Cell Tropism ◽  
Visna Virus ◽  
Pcr Products ◽  
The Central Nervous System ◽  
Maedi Visna Virus

Maedi-visna virus (MVV) is a lentivirus of sheep, mainly affecting the lungs and the central nervous system. Long terminal repeat (LTR) sequence variability is common in tissue culture-derived isolates of MVV as well as those of other lentiviruses. The role of this sequence variation in MVV replication has not been explored. PCR amplification of the LTRs of an MVV isolate revealed two product sizes, the larger containing a 53 bp duplication. PCR products containing the two size variants of the LTRs were cloned into an infectious molecular clone of MVV and the resulting chimeric viruses were tested for growth in various cell types. The chimeric virus containing only one copy of the 53 bp sequence was found to grow more slowly in sheep choroid plexus cells, sheep fibroblasts and sheep synovial cells than the virus with the 53 bp duplication. Both viruses grew equally well in macrophages. These results indicate that the LTRs determined the extended cell tropism of MVV.

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