polymerase chain reaction method
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2022 ◽  
Vol 15 (6) ◽  
pp. 669-684
G. B. Dikke ◽  
V. V. Ostromenskii ◽  
Yu. G. Kucheryavaya

Aim: to evaluate the efficacy of azoximer bromide along with a combined antimicrobial drug in treatment of patients with aerobic vaginitis (AV).Materials and Methods. There was conducted a prospective, randomized, parallel group study with enrolled 60 patients diagnosed with AV and 10 apparently healthy women (control group III). Group I (n = 30) received azoximer bromide (10 vaginal insertion according to the scheme, course of 10 days) and a combined antimicrobial drug, while Group II (n = 30) received only antimicrobial drug (course of 10 days). Microscopy of vaginal discharge smears, polymerase chain reaction method was used, detection of cytokines – interleukins (IL) and tumor necrosis factor-alpha (TNF-α) in vaginal washings was performed by ELISA. The study of neutrophil extracellular traps (NETs) was performed by ELISA, and DNA-containing strands ejected by neutrophils were determined by fluorescence method.Results. Recovery from disease was found in 100 % (30/30) and 86.7 % (26/30) in group I and group II, respectively. After treatment, serum level of IL-8 was revealed to decline in group I from 35.2 to 5.5 pg/ml (p = 0.05), in group II – from 33.4 to 5.3 pg/ml (p = 0.04), also not differ (p > 0.05) from the control values (5.2 pg/ml). The level of IL-1β also decreased after treatment in group I from 51.5 to 15.1 pg/ml (p = 0.002), in group II – from 57.9 to 20.1 pg/ml (p = 0.03), which also did not differ (p > 0.05) from the control values (16.7 pg/ml). The IL-10 level in both main groups decreased slightly (from 0.26 and 0.24 to 0.16 pg/ml in both groups), which was higher by 1.8-fold than in the control group (0.09 pg/ml; p < 0.001). The number of ejected NETs in both main groups decreased significantly after treatment, reaching control level. No side effects were observed, compliance and acceptability were 100 % in both groups. The frequency of AV relapses within 3 months was observed in 3.3 % (1/30) and 14.3 % (4/28) patients, after 6 months – in 14.3 % (4/28) and in 20.0 % (5/25) patients in groups I and II, respectively. Using the logit regression model, demonstrated that AV relapse might occur with a 50 %-probability at TNF-α level < 0.1 pg/ml after treatment (sensitivity – 67 %, specificity – 79 %). The risk of relapses in group I, in contrast to group II, showed a downward trend.Conclusions. The use of azoximer bromide and a combined broad-spectrum antimicrobial drug is effective in AV treatment, exerts a modulating effect on immune response parameters contributes to lowering relapse rate.

2022 ◽  
Vol 8 (12) ◽  
pp. 430-432
Chirantap Oza ◽  
Madhura Karguppikar ◽  
Vaman Khadilkar ◽  
Anuradha Khadilkar

Autoimmune polyglandular syndrome-1 (APS-1)also known as autoimmune polyendocrinopathy candidiasis ectodermal dystrophy is a rare autosomal recessive disorder caused by mutation of AIRE gene on chromosome 21q22.3 with an overall prevalence of <1:100,000. Here, we present a 16-year-old male having clinical history of evolution of symptoms for oral candidiasis, hypoparathyroidism, and adrenal insufficiency (AI). He developed rare endocrine and non-endocrine manifestations such as type-1 diabetes (T1D) and autoimmune hepatitis, respectively. The patient while on hormone replacement therapy along with immunosuppressants developed liver cirrhosis and portal hypertension with esophageal varices and candidiasis. Subsequently, he was admitted for complaints of cough, cold and fever and was confirmed to be affected by SARS-CoV-2 by reverse transcription-polymerase chain reaction method. In his prolonged ICU stay of 26 days, he required oxygen therapy, intravenous glucocorticoids, remdesivir, low molecular weight heparin, and hemodynamic support with inotropes. His medical management with subcutaneous insulin therapy and azathioprine was continued. He was discharged after complete resolution of symptoms and negative tests for SARS-CoV-2 and was advised radiological and clinical follow-up. Reports suggest that risk of severe COVID does not increase in patients with AI or autoimmunity. However, our patient possibly developed severe COVID not only due to AI and autoimmunity but also associated rare manifestations like hyperglycemia due to T1D and cirrhosis. Thus, good glycemic control and well-tolerated modern immunosuppressant therapy may be useful in improving prognosis of severe COVID-19 illness in patients with APS-1.

2021 ◽  
Vol 2131 (2) ◽  
pp. 022061
I G Kiselev ◽  
D A Belov

Abstract An urgent task is the development of devices that provide a precision temperature change. The devices used in the real-time polymerase chain reaction method require not only repeated cyclic temperature changes with high accuracy, but also the temperature gradient minimization over the working fluid. In this article, the modeling of the working processes taking place in the thermal block of the device with the previously proposed thermohydraulic system, which ensures the temperature change together with the Peltier elements, is carried out. A mathematical model is formulated and a differential equation, boundary and initial conditions are substantiated. Numerical methods for solving the mathematical model in the Elcut software were applied. An experimental setup was developed and a study of heat exchange processes in the thermal block of a nucleic acid analyzer with a thermohydraulic system was carried out, confirming the adequacy of the developed mathematical model.

2021 ◽  
Vol 25 (8) ◽  
pp. 1409-1414
O.C. Adekunle ◽  
O.S. Bolaji ◽  
A.O. Olalekan ◽  
T.O. Oyakeye

Infections associated with Staphylococcus aureus (S.aureus) have high mortality rate and lead to economic loss with a long stay in the hospital. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major nosocomial pathogens which are acquired in the health care facilities. The objective of the study is to investigate the presence of methicillin-resistant Staphylococcus aureus (MRSA) in clinical sources and hospital environments. Samples were collected, cultured and identified morphologically. Likewise, the antibiotic susceptibility profile was done. Identification was also done molecularly using PCR (Polymerase Chain Reaction) method. Sixty-three isolates were positive for S. aureus out of 370 clinical samples (urine, wound, nasal swabs and pus) and 37 positives out of 262 samples from hospital environments. Majority of the isolates were sensitive to cefoxitin, novobiocin and majority were resistant to ceftazidine, cloxacillin and augmentin. Seventy-one to ninety- five percent and 51-72% exhibited multi-drug resistance among clinical samples and hospital environments respectively. Both samples were positive for nuc and mec A genes. The detection of methicillin-resistant S. aureus (MRSA) in hospital environments may pose a great danger to patients especially those of compromised status.

2021 ◽  
pp. 3015-3020
Chommanad Lerdkrai ◽  
Nuch Phungphosop

Background and Aim: A canine multi-drug resistance 1 (MDR1) nt230(del4) is a well-known inherited disorder that primarily affects collies and various herding breeds. The most recognized clinical implication for affected dogs is associated with an increased risk of multiple drug toxicity. To date, MDR1 gene mutations have been identified globally, especially in dogs from the USA and European countries. This study aimed to investigate the prevalence of MDR1 nt230(del4) in herding dog breeds and Thai Ridgebacks in Thailand. Materials and Methods: We clarified the prevalence of MDR1 nt230(del4) in 263 dogs of eight purebred dog breeds in Thailand using an allele-specific multiplex polymerase chain reaction method and direct DNA sequencing. Results: Rough Collies, Australian Shepherds, Shetland Sheepdogs, and Old English Sheepdogs were affected by the mutation with mutant allelic frequencies of 57.14%, 12.82%, 11.28%, and 8.33%, respectively. Among these populations, the prevalence of the MDR1 (+/–) genotype was 57.14% (12/21) for Rough Collies, 25.64% (10/39) for Australian Shepherds, 16.13% (15/93) for Shetland Sheepdogs, and 16.67% (2/12) for Old English Sheepdogs, whereas the MDR1 (–/–) mutation was only identified in Rough Collies and Shetland Sheepdogs, with prevalences of 28.57% (6/21) and 3.22% (3/93), respectively. However, the MDR1 nt230(del4) was not identified in Border Collies, German Shepherds, White Swiss Shepherds, or Thai Ridgebacks. Conclusion: This study provides the current situation regarding MDR1 nt230(del4) in herding dog breeds in Thailand. In this survey, we investigated for the first time the status of MDR1 genotype in Thai Ridgebacks. These results are helpful for veterinarians managing effective therapeutic plans for commonly affected dog breeds, and these results will encourage all breeders to improve their selective breeding programs based on the MDR1 nt230(del4) status.

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2239
Zhen Liu ◽  
Zhenfei Dong ◽  
Binhui Zhan ◽  
Shifang Li

Apple (Malus domestica) fruits exhibiting bright stripe symptoms were identified in Weihai City, Shandong Province, China. To investigate the virome in the apple samples, the method of high throughput sequencing (HTS) was used to identify the viruses. It was found that the sequence of citrus concave gum-associated virus (CCGaV) was involved in the apple transcriptome dataset. The full-length genome of the CCGaV-Weihai isolate contained two segments, the RNA1 was 6674 nt in size containing a conserved RNA-dependent RNA polymerase (RdRp), and the RNA2 was ambisense, 2706 nt in length, encoding a movement protein (MP) and a coat protein (CP). Sequence alignment and phylogenetic analyses indicated that CCGaV-Weihai was more closely related to CCGaV-H2799 isolated from the apple host in the United States and distantly related to CCGaV-CGW2 from Citrus sinensis in Italy, indicating a possibly geographical and host differentiation of CCGaV isolates. This was the first identification and characterization of CCGaV infecting apples in China. Additionally, a rapid and sensitive reverse transcription recombinase polymerase amplification (RT-RPA) assay technique was established for CCGaV detection in apple plants. The RT-RPA of CCGaV was not affected by other common viruses in apple plants and is about 10-fold more sensitive than the conventional reverse transcription polymerase chain reaction method, which can be used in large-scale testing.

2021 ◽  
Vol 55 (4) ◽  
pp. 568-579
Tuğçe Şimşek Bozok ◽  
Ferit Kuşcu ◽  
Taylan Bozok ◽  
Süheyla Kömür ◽  
Aslıhan Candevir ◽  

2021 ◽  
Vol 13 (2) ◽  
pp. 181
Nadia Ayuningthias ◽  
Hasan Nasrullah ◽  
Dinar Tri Soelistiyowati ◽  
Eni Kusrini ◽  
Alimuddin Alimuddin

Highlight ResearchThe F2 of GH-transgenic B. imbellis was successfully producedThe transgene inheritance by the F2 fish was more than 90%The growth and body size of transgenic fish was significantly higher than controlF2 fish reached a larger body size in a shorter period compared to the F1 AbstractIn our previous research, we had successfully produced G0 and G1 Pangasianodon hypophthalmus growth hormone (PhGH) transgenic B. imbellis, native ornamental betta from Indonesia, which its giant-sized variant has valuable price for the breeders. The G0 and G1 transgenic (TG) fish showed higher growth rate and body size compared to the non-transgenic (NT) fish. The study was aimed to produce and evaluate the consistencies of transgene transmission and expression in G2 generation. The growth rate and body size between TG and NT fish was also compared. The G2 generation was produced using crosses between TG and NT G1 fish: ♂TG × ♀TG, ♂TG × ♀NT, ♂NT × ♀TG, and ♂NT ×♀ NT. Fish were reared for 12 weeks, and transgene detection was performed using the polymerase chain reaction method (PCR) on isolated DNA from the caudal fin clips. The endogenous and exogenous GH expression analysis was conducted using the quantitative real-time PCR (qPCR) method. The results showed that the inheritance of the GH transgene by the G2 fish was more than 90% in all transgenic crosses. Endogenous GH was expressed at the same levels in the brain of TG and NT fish, but the exogenous GH expression was highly detected only in the TG fish. The G2 transgenic fish had a higher specific growth rate, up to 31%, compared to the control. The body length of TG crosses were 23−35% higher and had 111−135% higher body weight compared to NT fish. These results showed a promising approached in mass-producing stable lines of giant-sized betta using the GH-transgenic technology.

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