scholarly journals Detecting the presence of bacterial RNA by polymerase chain reaction in low volumes of preoperatively aspirated synovial fluid from prosthetic joint infections

2020 ◽  
Vol 9 (5) ◽  
pp. 219-224
Author(s):  
B. Yang ◽  
X. Fang ◽  
Y. Cai ◽  
Z. Yu ◽  
W. Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Revision Surgery ◽  
Predictive Value ◽  
Quantitative Polymerase Chain Reaction ◽  
Joint Fluid ◽  
Chain Reaction ◽  
Musculoskeletal Infection ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections ◽  
Polymerase Chain

Aims Preoperative diagnosis is important for revision surgery after prosthetic joint infection (PJI). The purpose of our study was to determine whether reverse transcription-quantitative polymerase chain reaction (RT-qPCR), which is used to detect bacterial ribosomal RNA (rRNA) preoperatively, can reveal PJI in low volumes of aspirated fluid. Methods We acquired joint fluid samples (JFSs) by preoperative aspiration from patients who were suspected of having a PJI and failed arthroplasty; patients with preoperative JFS volumes less than 5 ml were enrolled. RNA-based polymerase chain reaction (PCR) and bacterial culture were performed, and diagnostic efficiency was compared between the two methods.According to established Musculoskeletal Infection Society (MSIS) criteria, 21 of the 33 included patients were diagnosed with PJI. Results RNA-based PCR exhibited 57.1% sensitivity, 91.7% specificity, 69.7% accuracy, 92.3% positive predictive value, and 55.0% negative predictive value. The corresponding values for culture were 28.6%, 83.3%, 48.5%, 75.0%, and 40.0%, respectively. A significantly higher sensitivity was thus obtained with the PCR method versus the culture method. Conclusion In situations in which only a small JFS volume can be acquired, RNA-based PCR analysis increases the utility of preoperative puncture for patients who require revision surgery due to suspected PJI. Cite this article: Bone Joint Res. 2020;9(5):219–224.

scholarly journals Use of a new multiplex quantitative polymerase chain reaction based assay for simultaneous detection of Neisseria meningitidis, Escherichia coli K , Streptococcus agalactiae, and Streptococcus pneumoniae

2021 ◽  
Author(s):  
Nastaran Hemmati ◽  
Farhad Nikkhahi ◽  
Amir Javadi ◽  
Sahar Eskandarion ◽  
Seyed Mahmoud Amin Marashi
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Streptococcus Pneumoniae ◽  
Bacterial Meningitis ◽  
Neisseria Meningitidis ◽  
Streptococcus Agalactiae ◽  
Predictive Value ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain

Background and Objectives: Neisseria meningitidis, Escherichia coli K , Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complica- tions. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. Materials and Methods: The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. Results: The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. Conclusion: The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

scholarly journals The use of polymerase chain reaction in the diagnosis and management of prosthetic joint infections: a debatable issues at this time

2019 ◽  
Vol 5 (2) ◽  
pp. 362
Author(s):  
Temi Ogunleye ◽  
Marlina Ponce de Leon ◽  
Suresh J. Antony
Keyword(s):  
Polymerase Chain Reaction ◽  
High Sensitivity ◽  
Chain Reaction ◽  
Diagnosis And Management ◽  
Joint Replacement Surgery ◽  
Replacement Surgery ◽  
Joint Infections ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections ◽  
Polymerase Chain

<p class="abstract">Joint replacement surgery is increasing due to its success in decreasing pain and restoring function. Prosthetic joint infections (PJI) is one of the most detrimental complications of the surgery. These infections can either be acute or chronic and can be caused by a variety of organisms. Effective and efficient identification of the cause of infection is vital so that proper treatment can be provided. The use of polymerase chain reaction (PCR) is a possibility for diagnosis and management of PJI with a reduction in the use of incorrect antibiotics. This is due to its ability to quickly diagnosis viral, bacterial, rickettsia, mycobacterial, and protozoal infection in hours. It also has high sensitivity and specificity even with antimicrobial usage and biofilm production. However, more studies need to be done in order to be able to classify it as a possible gold standard.</p>

scholarly journals Are There Benefits In Early Diagnosis Of Prosthetic Joint Infection With Multiplex Polymerase Chain Reaction?

2017 ◽  
Vol 2 (4) ◽  
pp. 175-183 ◽  
Author(s):  
Christian Lausmann ◽  
Akos Zahar ◽  
Mustafa Citak ◽  
Julian Brañes ◽  
Stefan Schmidl ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Early Diagnosis ◽  
Multiplex Pcr ◽  
Dna Sequences ◽  
Predictive Value ◽  
Joint Infection ◽  
Negative Control ◽  
Chain Reaction ◽  
Prosthetic Joint ◽  
Polymerase Chain

Abstract. Purpose Identification of bacteria and susceptibility are fundamental in periprosthetic joint infection (PJI). Especially in the case of systemic inflammatory response syndrome (SIRS) rapid detection of pathogens is essential for proper therapy. Bacterial cultures are time consuming. The polymerase chain reaction (PCR) is a non-culture molecular method and is able to rapidly identify pathogens and their resistance genes. Multiplex PCR (mPCR) can amplify several different DNA sequences simultaneously. The aim of this study was to show the value of mPCR for early diagnosis of PJI.Methods 60 patients undergoing total hip or knee revisions were recruited in this prospective single-centre-study. Three groups were created: 26 patients with aseptic loosening (negative control), 26 patients with chronic PJI, and 8 patients with acute PJI/SIRS. We compared the results of joint aspirates obtained intraoperatively investigated by mPCR with the microbiology results of tissue specimens.Results The overall sensitivity of mPCR was 78.8% (95% CI, 61.1 - 91.0%), the specificity was 100% (95% CI, 87.2 - 100%), the negative predictive value was 79.4% (95% CI, 62.1 - 91.3%), the positive predictive value was 100% (95% CI, 86.8 - 100%), and the overall accuracy was 88.3% (95% CI, 77.4 - 95.2%). The overall accuracy in acute infections/SIRS (87.5%) was greater than in late chronic PJI (76.9%). In PJI the mPCR was able to provide the results within 5 hours whereas the mean time for cultures was 6.4 days.Conclusions Multiplex PCR is a reliable diagnostic tool in PJI management, especially in acute cases complicated with SIRS. Early diagnosis within several hours is possible, targeted antibiotic treatment can be started promptly.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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