Sample Biological Material Operating Procedure for the method of molecular-biological techniques (PCR diagnostics)

2016 ◽  
Vol 3 (3) ◽  
pp. 156-160
Author(s):  
Valery A. Kaptilnyy

Review describes a method of diagnostics with the use of the polymerase chain reaction (PCR diagnostics) of infections in patients in modern obstetric-gynecological practice. Particular attention is paid to urogenital tract pathogens to be diagnosed by the method of nucleic acid amplification. There are described the preparation for the study, purposes, the technology of amplification procedure. There is shown the difference in the detection of the DNA and RNA pathogen. The presented photographs show in detail all the stages of obtaining scrapings from the urogenital tract.

1998 ◽  
pp. 414-436 ◽  
Author(s):  
Françoise Mathieu-Daudé ◽  
Karen Evans ◽  
Frank Kullmann ◽  
Rhonda Honeycutt ◽  
Thomas Vogt ◽  
...  

2019 ◽  
Vol 3 (2) ◽  
pp. 83
Author(s):  
Anita Kurniati ◽  
Desak Nyoman Surya Suameitra Dewi ◽  
Ni Nyoman Purwani

Background: Tuberculosis (TB) is one of the major causes of health burden worldwide, especially in lower middle-income countries. TB is caused by Mycobacterium tuberculosis (MTB) and characterized by severe condition incuding coughing and fever. Purpose: To review the current methods for detection of TB using Polymerase Chain Reaction (PCR). Review: several studies have been done to give valuable insight into TB transmission, diagnosis, and treatment, however research  is constantly  needed  to decrease the incidence of eradicate TB. This infectious disease still give big health problem in all over the world by being second in causing high mortality rates after HIV/AIDS.  A specific, sensitive, rapid and cheap method for TB and other mycobacteria diagnosis in clinical specimen is a desperate needed in the laboratory diagnosis and hence management of tuberculosis. PCR as one of nucleic acid amplification assays have revolutionized MTB detection. Since it was first invented in fifteen years ago, it’s been through many developments. Conclusion: PCR  is one of the most specific and sensitive method currently available for TB diagnosis that can also detect in in all types of specimens obtained from TB patients.


2019 ◽  
Vol 11 (43) ◽  
pp. 5568-5576
Author(s):  
Sonja Elf ◽  
Kevin E. Eboigbodin

SIBA is an established nucleic acid amplification method that is used as an alternative to polymerase chain reaction (PCR).


2017 ◽  
Vol 49 (8) ◽  
pp. 625-627
Author(s):  
Fumio Terasaki ◽  
Hitomi Fukumoto ◽  
Ryo Kawata ◽  
Yoshinobu Hirose ◽  
Shu-ichi Fujita ◽  
...  

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
John Mokua Mose ◽  
John Maina Kagira ◽  
Simon Muturi Karanja ◽  
Maina Ngotho ◽  
David Muchina Kamau ◽  
...  

The detection ofToxoplasma gondiiin free-range chickens is a good indicator of possible risk to human beings. The aim of this study was to investigate the occurrence ofT. gondiiin free-range chicken using polymerase chain reaction (PCR). Brain samples from 105 free-range chickens from three administrative areas in Thika region, Kenya, were collected, DNA-extracted, and analyzed using PCR to detect presence ofT. gondii. The overall prevalence ofT. gondiiin all the three areas was 79.0% (95% CI: 70.0–86.4%) and the prevalence across the three areas was not significantly different (P=0.5088;χ2=1.354). Female chickens had higher (79.4%) prevalence than males (78.6%), although the difference was not significant (P=0.922,χ2= 0.01). However, chickens that were more than 2 years old had significantly (P=0.003;χ2= 11.87) higher prevalence compared to younger ones. The study indicates that there was a high occurrence ofT. gondiiinfection in free-range chickens from Thika region and that the infection rate is age dependent. Further studies should be carried out to determine the possible role of roaming chickens in the epidemiology of the disease among humans in the area.


2020 ◽  
Vol 21 (1) ◽  
pp. 42-48
Author(s):  
D. G. Pochernikov ◽  
V. V. Getman ◽  
N. T. Postovoytenko ◽  
D. M. Rysev ◽  
I. S. Galkina

The study objective is to compare the rate of detection of various microorganisms in the prostatic fluid and ejaculate using real-time polymerase chain reaction in patients with category IV chronic prostatitis.Materials and methods. Between December of 2016 and July 2019, a prospective study including 81 patients with category IV chronic prostatitis per the National Institutes of Health Prostatitis Syndrome Classification (1999) was performed. The patients referred to the clinic of the Ivanovo State Medical Academy for preconception preparation, infertility or erectile disfunction. At the examination, all patients lacked symptoms characteristic of category II or III chronic prostatitis. Transrectal ultrasound of the prostate, microscopic examination of the prostatic fluid and (or) ejaculate, quantitative examination of urogenital tract microbiota using real-time polymerase chain reaction were performed.Results. Comparison of microbiota of the prostatic fluid and ejaculate showed significant differences in the total amount of bacterial mass: in the prostatic fluid mean titer was 3.7 ± 1.6, in the ejaculate it was 2.6 ± 1.8 (p <0.001). Prostatic fluid contained significantly more of the following microorganisms: Enterobacteriaceae spp./Enterococcus spp., Staphylococcus spp., Streptococcus spp., Corynebacterium, Eubacterium, Anaerococcus (p <0.05). No significant differences in the amounts of other microorganisms were observed.Conclusion. The study demonstrates significant heterogeneity of qualitative and quantitative microbiota content in the prostatic fluid and ejaculate. Supposedly, it can be explained by anatomical and physiological characteristics of the prostate, seminal vesicles and periurethral glands that secrete fluid for the ejaculate. The ejaculate contains less microorganisms compared to prostatic fluid which should be taken into account in differential diagnosis of infections of the urogenital tract.The authors declare no conflict of interest.All patients gave written informed consent to participate in the study.


2008 ◽  

The book "La PCR e le sue varianti" is designed as a reference tool for those whose laboratory activities deal with methods based on nucleic acid amplification. The text provides the theoretical bases of the polymerase chain reaction (PCR) and its variants (e.g. RT-PCR, quantitative PCR, isothermic PCR) in a rapid and concise manner and describes the principal applications used for genetic identification and the study of genetic polymorphism, in the form of a protocol that can be easily consulted by the users.


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