Successful containment of horizontal enterovirus infection in a neonatal unit in Singapore through diagnosis by polymerase chain reaction (PCR) and direct sequence analysis
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Successful containment of horizontal enterovirus infection in a neonatal unit in Singapore through diagnosis by polymerase chain reaction (PCR) and direct sequence analysis
Background: Enterovirus’ epidemics (EV) often coincide with seasonal peaks in the community. However, they can also occur sporadically in neonatal units. The identification of EV infection in newborns can be difficult because they tend to have light or non-specific symptoms. This study reports an EV epidemic in the neonatal unit of KK Women’s Hospital, Singapore.
Methods: This is a simple and retrospective retrospective cohort study of newborns that had positive results for EV during the epidemic. Demographic characteristics, clinical presentations and results have been analyzed. The control measures used to limit the propagation of the infection are reported.
Results: A total of 7 cases of EV infection have been identified. Their median weighing on birth and their gestational age were 1240 g (750 to 2890 g) and 28 weeks (26-35 weeks), respectively. The symptoms took place at a median age of 48 days (9-103 days). All cases initially presented with recurrent apnea and 4 necessary for the assisted fan required with CPAP (2) and mechanical ventilation (2). Serious complications have occurred in 3 infants (2 with a necroting enterocolite and 1 with meningitis) and no deceased. EV has been detected from the relevant swabs (n = 6), csf (n = 2) and nasopharyngeal (n = 2). The viral subtyping revealed an Echovirus 25. The monitoring of all infants exposed by nasopharyngeal swabs was implemented, as well as strict contact precautions and coherence measures.
Conclusions: Premature infants with EV are more subject to serious complications, which can lead to significant morbidity. Thus, early recognition of symptoms, rapid diagnosis and rapid implementation of infection control measures are essential to prevent propagation of subsequent infection.
Kallikreins of human tissue in polymorphic adenocarcinoma: a polymerase chain reaction and an immunohistochemical study
The polymorphic adenocarcinoma (PAC) is the second most common tumor of malignant salivary gland of minor salivary glands. The Kallikreins (KLKS) human tissues are a very preserved family of proteases expressed by various tissues and organs. The literature demonstrates a link between klks and the salivary neoplasms of the gland. The purpose of this study was to determine KLK mRNA levels in CAP tissue samples and determine whether the KLK expression is limited to tumor cells. Nineteen CAP cases were examined (1987-2013). The diagnosis has been confirmed, demographic data has been collected and samples of formulated fixed salivary tissues and fixed paraffin. RNA insulation has been achieved, followed by complementary DNA conversion through reverse transcription. By using PCR, the quantitative level of expression of KLKS1-15 has been recorded.
Samples with high and low kLK expression were selected for the coloring of immunohistochemistry. The results revealed a statistically significant increase in KLK average KLK mRNA expression for KLK1, KLK4, KLK10, KLK12 and KLK15 in PAC tissue samples, compared to normal salivary gland fabric (U Mann-Whitney U test, P <0.05). The results of immunohistochemistry have shown that KLKS was present in tumor cells. In particular, all samples demonstrating a relatively higher KLK mRNA expression showed equivalent or increased staining scores compared to low KLK mRNA expression samples. In conclusion, there appears to be an aberrant kallikrein expression in polymorphic adenocarcinoma, suggesting that the possibility of influence on Kallikrein’s waterfall on the development and progression of the tumor. Opportunine identification of etiological agents of enteric infections is necessary to reduce the burden of infantile diarrheal mortality.
The detection methods based on the amplification of the nucleic acid provide a fast and reliable way for the diagnosis of microbes in clinical samples. This study was undertaken to evaluate a conventional conventional polymeric chain reaction of conventional conventional transcription (RT-PCR) conventional (RT-PCR) transcription “developed at the Indian Council for Medical Research.
Successful containment of horizontal enterovirus infection in a neonatal unit in Singapore through diagnosis by polymerase chain reaction (PCR) and direct sequence analysis
Double Disclosure Probes Improve the sensitivity of detection to acute respiratory syndrome Coronavirus 2 (SARS-COV-2) in a reaction of the reverse transcription polymerase chain (RT-PCR)
Background: Serious Respiratory Syndrome Coronavirus 2 (SARS-COV-2) that emerged in the city of Wuhan, Province of Hubei, China, spread all over the world and threatens human life. The detection of SARS-COV-2 is essential to prevent new epidemics, the retrocedure of the distribution of diseases and the management of patients. Currently, a reaction of the reverse transcription polymerase chain (RT-PCR) is used to detect the virus in clinical laboratories. However, although this test is considered a high specificity, its sensitivity would be as low as 60-70%. Improved sensitivity is therefore urgently required.
Methods: We used the first-handed program and a range recommended by the CDC (N1, N2 and N3) in the United States and NIID (N1 and N2) in Japan. In addition, we have designed double mess probes according to the virus sequence provided by the NIID to develop another test (called the YCH test [N1 and N2]). Using these tests, we made RT-PCR with serial diluted DNA positive controls to evaluate and compare the detection sensitivity of the three tests. In addition, 66 nasopharyngeal swabs were tested to determine diagnostic performance.
Results: The value of the RT-PCR threshold cycle (CT) was relatively small for CDC and YCH analyzes in relation to the NIID test. Serial dilution tests have shown that CDC and YCH analyzes could detect low copy numbers of the positive control of DNA. The starting background fluorescence signal was lower for the YCH test compared to the NIID test. We evaluated the diagnostic performance between single probes (NIID) and double incorrect (YCH) using 66 nasopharyngeal buffers. When the results of the YCH-N2 analysis were used as a reference, each test has detected SARS-COV-2 with 56% positive percentage agreements for NIID-N1, 61% for YCH-N1 and 94% for NIID -N2, and 100% of negative agreements for NIID-N1, YCH-N1 and NIID-N2.
Description: DNA polymerase eta (POLH), is a protein that in humans is encoded by the POLH gene. This gene encodes a member of the Y family of specialized DNA polymerases. It copies undamaged DNA with a lower fidelity than other DNA-directed polymerases. However, it accurately replicates UV-damaged DNA; when thymine dimers are present, this polymerase inserts the complementary nucleotides in the newly synthesized DNA, thereby bypassing the lesion and suppressing the mutagenic effect of UV-induced DNA damage. This polymerase is thought to be involved in hypermutation during immunoglobulin class switch recombination. Mutations in this gene result in XPV, a variant type of xeroderma pigmentosum. Several transcript variants encoding different isoforms have been found for this gene.
Description: DNA polymerase iota is an enzyme that in humans is encoded by the POLI gene. The protein encoded by this gene is an error-prone DNA polymerase involved in DNA repair. The encoded protein promotes DNA synthesis across lesions in the template DNA, which other polymerases cannot do. The encoded polymerase inserts deoxynucleotides across lesions and then relies on DNA polymerase zeta to extend the nascent DNA strand to bypass the lesion.
Description: The protein encoded by this gene is a DNA polymerase involved in base excision and repair, also called gap-filling DNA synthesis. The encoded protein, acting as a monomer, is normally found in the cytoplasm, but it translocates to the nucleus upon DNA damage. [RefSeq]
Description: Polymerase (DNA directed), beta, also known as POLB, is an enzyme that, in humans, is encoded by the POLB gene. It is localized on 8p11.2. The protein encoded by this gene is a DNA polymerase involved in base excision and repair, also called gap-filling DNA synthesis. It is found that a truncated POLB is expressed in primary colorectal tumors and inhibits the normal repair function of wildtype POLB. The encoded protein, acting as a monomer, is normally found in the cytoplasm, but it translocates to the nucleus upon DNA damage. Several transcript variants of this gene exist, but the full-length nature of only one has been described to date. Additionally, human POLB forms a complex with and is methylated by PRMT6. In vitro, methylated POLB possesses significantly higher DNA polymerase activity when compared to that of unmodified enzyme. The increase in DNA polymerase activity upon methylation is due to the enhanced DNA binding and processivity of POLB.
Description: DNA polymerase lambda, also known as POLL, is a protein found in eukaryotes. In humans, it is encoded by the POLLA gene. POLL has 5-prime-deoxyribose-5-phosphate lyase activity and strand-displacement synthesis activity on gapped DNA substrates, suggesting that POLL participates in short- and long-patch base excision repair. POLL is widely expressed, with abundant expression in pachytene spermatocytes of testis and in ovary.
