scholarly journals A Novel Microfluidic Device Integrated with Chitosan-Modified Capillaries for Rapid ZIKV Detection

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 186 ◽  
Author(s):  
Xinchao Zhu ◽  
Jun Zhao ◽  
Anzhong Hu ◽  
Jingyu Pan ◽  
Guoqing Deng ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Silicon Dioxide ◽  
Microfluidic Chip ◽  
Polymerase Chain Reaction Amplification ◽  
Signal Acquisition ◽  
Acid Extraction ◽  
Chain Reaction ◽  
In Situ Pcr ◽  
Polymerase Chain

The outbreak of Zika virus (ZIKV) has posed a great challenge to public health in recent years. To address the urgent need of ZIKV RNA assays, we integrate the microfluidic chip embedded with chitosan-modified silicon dioxide capillaries, smartphone-based detection unit to be a C3-system for the rapid extraction and detection of ZIKV RNA. The C3-system is characterized by: (1) four chitosan-modified silicon dioxide capillaries integrated in the microfluidic chip for target ZIKV RNA enrichment and “in situ PCR” (polymerase chain reaction) amplification; (2) smartphone-based point of care (POC) device consisting of a pneumatic subsystem for controlling the nucleic acid extraction processes in the microfluidic chip, a heating subsystem for sample lysis and PCR amplification, and an optical subsystem for signal acquisition. The entire detection processes including sample lysis, ZIKV RNA enrichment, and reverse-transcription polymerase chain reaction (RT-PCR) is achieved in the microfluidic chip. Moreover, PCR buffers can be directly loaded into the chitosan-modified silicon dioxide capillaries for “in situ PCR”, in which the captured ZIKV RNA is directly used for downstream PCR without any loss. ZIKV RNA extracted by the C3-system can be successfully recovered at very low concentrations of 50 transducing units (TU)/mL from crude human saliva. This means that our method of detecting viremia in patients infected with ZIKV is reliable.

Enterovirus is Not Present in Placentas from Cases of Perinatal Depression Using Polymerase Chain Reaction Analysis

2009 ◽  
Vol 12 (3) ◽  
pp. 177-179 ◽  
Author(s):  
Jesse Cover ◽  
Edwina Popek ◽  
Myra Wyckoff ◽  
N. Kristine Leos ◽  
Beverly Barton Rogers
Keyword(s):  
Polymerase Chain Reaction ◽  
Respiratory Insufficiency ◽  
Pcr Amplification ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Apgar Scores ◽  
Specific Patient ◽  
In Situ Pcr ◽  
Polymerase Chain

Enteroviruses have been implicated as a cause of low Apgar scores in conjunction with perinatal seizures and respiratory insufficiency. Using in-situ reverse transcriptase polymerase chain reaction (in-situ PCR), Nuovo et al detected enterovirus in up to 86% of placentas from perinates exhibiting these symptoms. In-situ PCR has been the only method employed to assess for the presence of enterovirus in this specific patient population. The purpose of our study was to use PCR amplification of enterovirus from extracted RNA to confirm these observations. RNA was extracted from 26 placentas of infants with low Apgar scores, perinatal seizures, and respiratory insufficiency. Each extraction was positive for beta-actin RNA, which confirmed that the integrity of RNA was maintained in the sample. Enterovirus RNA was not detected in any of the cases. Our results indicate that enterovirus is not present in placentas from neonates with the combination of low Apgar scores, respiratory insufficiency, and seizures, as previously reported.

In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue

1998 ◽  
Vol 70 (2) ◽  
pp. 119-127 ◽  
Author(s):  
Pádraig M Strappe ◽  
Ting Huei Wang ◽  
Chris-Anne McKenzie ◽  
Suzanne Lowrie ◽  
Peter Simmonds ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Brain Tissue ◽  
Polymerase Chain Reaction Amplification ◽  
Chain Reaction ◽  
Reaction Amplification ◽  
Polymerase Chain ◽  
Hiv 1

scholarly journals The In Situ Polymerase Chain Reaction for Detection of Chlamydia trachomatis

1998 ◽  
Vol 46 (9) ◽  
pp. 1017-1023 ◽  
Author(s):  
Thilo Schlott ◽  
Götz Ruda ◽  
Michael Hoppert ◽  
Holger Nagel ◽  
Silke Reimer ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Cervical Neoplasms ◽  
Chain Reaction ◽  
Pcr Assay ◽  
In Situ Pcr ◽  
Polymerase Chain

The in situ polymerase chain reaction (PCR) is a technique that has important applications in the diagnosis of viral and bacterial diseases. This study investigated an in situ PCR assay established to detect the presence of Chlamydia trachomatis in endocervical swabs. In addition, histological sections of endocervical squamous cell carcinoma were analyzed because previous studies had revealed a significant association with C. trachomatis. A total of 20 cervical neoplasms (squamous cell carcinoma in situ; n = 10; invasive squamous cell carcinoma; n = 10) and endocervical smears taken from five patients with and without inflammatory changes were analyzed by conventional PCR. Chlamydial DNA was found in 10 histological samples (six carcinomas in situ, four invasive carcinomas) and in one endocervical swab from a patient with known C. trachomatis infection. Positive specimens were used for establishing an in situ PCR assay (IS-PCR). After IS-PCR, these samples showed dense cytoplasmic staining of endocervical cells (smears) and non-neoplastic epithelial cells (cervical neoplasms). The other tumor samples and smears did not demonstrate positive PCR reaction. The results indicate that in situ PCR is an effective technique for localizing C. trachomatis in target cells because IS-PCR detection of chlamydial DNA correlated with histological and cytological features.

scholarly journals Determination the causative strain for hydatid cyst in Iraqi cattle by using ND1 gene

2017 ◽  
Vol 41 (1) ◽  
pp. 11-16
Author(s):  
Mohammed J. Muhaidi
Keyword(s):  
Polymerase Chain Reaction ◽  
Nadh Dehydrogenase ◽  
Sequence Data ◽  
Polymerase Chain Reaction Amplification ◽  
Acid Extraction ◽  
Reaction Products ◽  
Hydatid Cysts ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sheep Strain

     Hydatid Cysts were obtained from 15 cows from liver, lung, spleen, heart, and peritoneal cavity, between December 2014 and October 2015. Hydatid cysts (protoscoleces) were used for deoxyribonucleic acid extraction by using mechanical grinder. The purification of mtDNA was done by (promega kit, USA). The mitochondrial NADH dehydrogenase subunit 1 (ND1) genes was used as targets for polymerase chain reaction amplification, all hydatid cysts yielded amplification products. Polymerase chain reaction product for NADH1 800 basic pair. The polymerase chain reaction products were purified and partial sequences were generated. The sequences obtained were found to align with corresponding region for ND1 gene in the Gene Bank nucleotide database confirming to genotype of sheep strain (G1) in Iraq, Phylogenetic analysis of partial sequence data from ND1 genes for obtained Phylogenetic tree. G1 genotype was the most common taxon and the actual source of infection of Iraqi's cattle. All of 15 strains were G1 genotype (sheep strain) based on the partial sequences of NADH dehydrogenase 1 (ND1).

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

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