Evaluation of Apparatus Used to Test Liquid through Protective Materials: Comparison of a Modified Dot-Blot Apparatus to the ASTM Penetration Cell

Abstract Effective personal protective equipment (PPE) is critical in preventing the spread of infectious diseases. Appropriate test systems and test soils are needed to adequately evaluate PPE. ASTM test method F903, which specifies the test method setup also used in ASTM F1670 and F1671, has been used for decades to test liquid (ASTM F1670) or viral (ASTM F1671) penetration resistance of PPE fabrics. However, an alteration of the bacteriophage propagation method detailed in the standard was necessary to obtain consistent titers of virus. In this study, modification of the nutrient broth provided consistently higher titers of virus and the use of the top agar in smaller increments prevented premature solidification. This study then compared the standard ASTM F1671 (using bacteriophage φχ174) with a modified dot-blot method to assess viral penetration of PPE materials. The results indicated that ASTM F1671 and the dot-blot apparatus methods were equivalent. The dot-blot method described here is less labor intensive and faster than the ASTM F1671 method. However, using the dot-blot system, which uses antibodies to detect the bacteriophage and signal amplification, does not indicate if virus viability or infectivity is retained, whereas the ASTM F1671 method indicates both. Nonetheless, the method presented in this investigation is a substantial improvement of a standard method for viral challenge testing of PPE materials.

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A combined ultrastructural and gene molecular research for the relationship between human uterine cervical carcinoma and human papillomavirus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010015856x ◽

1990 ◽

Vol 48 (3) ◽

pp. 204-205

Author(s):

Kun Lee ◽

Jingyi Si ◽

Ricai Han ◽

Wei Zhang ◽

Bingbing Tan ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Dot Blot ◽

Homologous Sequences ◽

Normal Cervical Epithelium ◽

The Relationship ◽

Chronic Cervicitis

There are more supports for the view that human papillomavirus (HPV) infection might be an etiological factor in the development of cervical cancer when the association of persistent condylomata is considered. Biopsies from 318 cases with squamous cell carcinoma of uterine cervix, 48 with cervical and vulvar condylomata, 14 with cervical intraepithelial neoplasia (CIN), 34 with chronic cervicitis and 24 normal cervical epithelium were collected from 5 geographic regions of China with different cervical cancer mortalities. All specimens were prepared for Dot blot, Southern blot and in situ DNA-DNA hybridizations by using HPV-11, 16, 18 DNA labelled with 32P and 3H as probes to detect viral homologous sequences in samples. Among them, 32 cases with cervical cancer, 27 with condyloma and 10 normal cervical epitheliums were randomly chosen for comparative EM observation. The results showed that: 1), 192 out of 318 (60.4%) cases of cervical cancer were positive for HPV-16 DNA probe (Table I)

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APPLICATION OF REAL-TIME PRC TECHNIQUE AND REVERSE DOT-BLOT FOR DETECTION THE HIGH RISK TYPES OF HUMAN PAPILLOMAVIRUS CAUSING CERVICAL CANCER

Journal of Medicine and Pharmacy ◽

10.34071/jmp.2017.3.15 ◽

2017 ◽

pp. 99-103

Author(s):

Van Bao Thang Phan ◽

Hoang Bach Nguyen ◽

Van Thanh Nguyen ◽

Thi Nhu Hoa Tran ◽

Viet Quynh Tram Ngo

Keyword(s):

Cervical Cancer ◽

High Risk ◽

Real Time ◽

Real Time Pcr ◽

Dot Blot ◽

The Real ◽

Dot Blot Assay ◽

Hpv Types ◽

High Risk Hpv ◽

Reverse Dot Blot

Introduction: Infection with HPV is the main cause of cervical cancer. Determining HPV infection and the types of HPV plays an important role in diagnosis, treatment and prognosis of cervicitis/cervical cancer. Aims: Determining proportion of high-risk HPV types and the occurrence of coinfection with multiple HPV types. Methods: 177 women with cervicitis or abnormal Pap smear result were enrolled in the study. Performing the real-time PCR for detecting HPV and the reverse DOT-BLOT assay for determining type of HPV in cases of positive PCR. Results: 7 types of high-risk HPV was dectected, the majority of these types were HPV type 18 (74.6%) and HPV type 16 (37.6%); the proportion of infection with only one type of HPV was 30.4% and coinfection with multiple HPV types was higher (69.6%), the coinfected cases with 2 and 3 types were dominated (32.2% and 20.3%, respectively) and the coinfected cases with 4 and 5 types were rare. Conclusion: Use of the real-time PCR and reverse DOT-BLOT assay can determine the high-risk HPV types and the occurrence of coinfection with multiple HPV types. Key words: HPV type, Reverse DOT-BLOT, real-time PCR,PCR, cervical cancer

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Detection of Hepatitis a Virus and other Enteroviruses in Wastewater and Surface Water Samples by Gene Probe Assay

Water Science & Technology ◽

10.2166/wst.1991.0071 ◽

1991 ◽

Vol 24 (2) ◽

pp. 267-272 ◽

Cited By ~ 12

Author(s):

S. Dubrou ◽

H. Kopecka ◽

J. M. Lopez Pila ◽

J. Maréchal ◽

J. Prévot

Keyword(s):

Surface Water ◽

Cell Cultures ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Limit Of Detection ◽

Blot Hybridization ◽

Noncoding Region ◽

Gene Probe ◽

Dot Blot ◽

Lower Efficiency

Enteroviruses were specifically detected by dot blot hybridization when using poliovirus type 1 (PV1) derived subgenomic radiolabeled cRNA probes (riboprobes) in environmental water specimens and in the cell cultures in which the viruses were amplificated. The riboprobe corresponding to the 5' noncoding sequence detected the majority of enteroviruses. Hepatitis A virus (HAV) was specifically detected by an HAV cRNA probe corresponding to the 5' noncoding region of its genome. By this test, the limit of detection of coxsackievirus B5 and echovirus 7 seeded in mineral water was 103 to 104 PFU/spot. In cell cultures, positive signals were observed in the lysates of cells infected by one PFU. Higher positive signals were obtained with a short PV1 probe (nt 221-670) corresponding to the 5' noncoding region, which is a well preserved sequence among the enteroviruses, than with PV1 genomic probe. Hybridization allowed a good detection of enteroviral RNAs in wastewater specimens, but with a lower efficiency in surface water. In this case, amplification of viruses in the cell cultures gave significant hybridization results.

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Feasibility of Qualitative Testing of BCR-ABL and JAK2 V617F in Suspected Myeloproliperative Neoplasm (MPN) Using RT-PCR Reversed Dot Blot Hybridization (RT-PCR RDB)

Clinical Lymphoma Myeloma & Leukemia ◽

10.1016/j.clml.2019.01.005 ◽

2019 ◽

Vol 19 (4) ◽

pp. 220-227

Author(s):

Najmiatul Masykura ◽

Ummu Habibah ◽

Siti Fatimah Selasih ◽

Soegiarto Gani ◽

Cosphiadi Irawan ◽

...

Keyword(s):

Blot Hybridization ◽

Jak2 V617f ◽

Rt Pcr ◽

Dot Blot ◽

Dot Blot Hybridization

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Use of monoclonal and polyclonal antibodies to detect prostatic acid phosphatase by immunoblotting.

Clinical Chemistry ◽

10.1093/clinchem/32.10.1832 ◽

1986 ◽

Vol 32 (10) ◽

pp. 1832-1835 ◽

Cited By ~ 2

Author(s):

P C Patel ◽

L Aubin ◽

J Côte

Keyword(s):

Monoclonal Antibodies ◽

Acid Phosphatase ◽

Western Blot ◽

Polyclonal Antibodies ◽

Prostatic Acid Phosphatase ◽

The Other ◽

Dot Blot ◽

Western Blots ◽

Dot Blot Assay ◽

Polyclonal Antisera

Abstract We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic protocols. We conclude that PAP can be readily detected by Western blots with use of polyclonal antisera, but not with monoclonal antibodies. On the other hand, using a dot blot assay, we could easily detect PAP with both polyclonal and monoclonal antibodies.

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A Novel Mouse Monoclonal Antibody C42 against C-Terminal Peptide of Alpha-1-Antitrypsin

International Journal of Molecular Sciences ◽

10.3390/ijms22042141 ◽

2021 ◽

Vol 22 (4) ◽

pp. 2141

Author(s):

Srinu Tumpara ◽

Elena Korenbaum ◽

Mark Kühnel ◽

Danny Jonigk ◽

Beata Olejnicka ◽

...

Keyword(s):

Monoclonal Antibody ◽

Flow Cytometry ◽

Western Blotting ◽

Complex Mixture ◽

Immunoglobulin M ◽

Confocal Laser ◽

Enzyme Linked Immunosorbent Assay ◽

Dot Blot

The C-terminal-fragments of alpha1-antitrypsin (AAT) have been identified and their diverse biological roles have been reported in vitro and in vivo. These findings prompted us to develop a monoclonal antibody that specifically recognizes C-36 peptide (corresponding to residues 359–394) resulting from the protease-associated cleavage of AAT. The C-36-targeting mouse monoclonal Immunoglobulin M (IgM) antibody (containing κ light chains, clone C42) was generated and enzyme-linked immunosorbent assay (ELISA)-tested by Davids Biotechnologie GmbH, Germany. Here, we addressed the effectiveness of the novel C42 antibody in different immunoassay formats, such as dot- and Western blotting, confocal laser microscopy, and flow cytometry. According to the dot-blot results, our novel C42 antibody detects the C-36 peptide at a range of 0.1–0.05 µg and shows no cross-reactivity with native, polymerized, or oxidized forms of full-length AAT, the AAT-elastase complex mixture, as well as with shorter C-terminal fragments of AAT. However, the C42 antibody does not detect denatured peptide in SDS-PAGE/Western blotting assays. On the other hand, our C42 antibody, unconjugated as well as conjugated to DyLight488 fluorophore, when applied for immunofluorescence microscopy and flow cytometry assays, specifically detected the C-36 peptide in human blood cells. Altogether, we demonstrate that our novel C42 antibody successfully recognizes the C-36 peptide of AAT in a number of immunoassays and has potential to become an important tool in AAT-related studies.

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Exogenous Isolation of Antibiotic Resistance Plasmids from Piggery Manure Slurries Reveals a High Prevalence and Diversity of IncQ-Like Plasmids

Applied and Environmental Microbiology ◽

10.1128/aem.66.11.4854-4862.2000 ◽

2000 ◽

Vol 66 (11) ◽

pp. 4854-4862 ◽

Cited By ~ 147

Author(s):

Kornelia Smalla ◽

Holger Heuer ◽

Antje Götz ◽

Dagmar Niemeyer ◽

Ellen Krögerrecklenfort ◽

...

Keyword(s):

Antibiotic Resistance ◽

Ralstonia Eutropha ◽

Blot Hybridization ◽

Restriction Pattern ◽

Pcr Analysis ◽

Dot Blot ◽

High Prevalence ◽

Resistance Plasmids ◽

Incq Plasmids ◽

K 12

ABSTRACT Antibiotic resistance plasmids were exogenously isolated in biparental matings with piggery manure bacteria as plasmid donors inEscherichia coli CV601 and Pseudomonas putidaUWC1 recipients. Surprisingly, IncQ-like plasmids were detected by dot blot hybridization with an IncQ oriV probe in severalP. putida UWC1 transconjugants. The capture of IncQ-like plasmids in biparental matings indicates not only their high prevalence in manure slurries but also the presence of efficiently mobilizing plasmids. In order to elucidate unusual hybridization data (weak or no hybridization with IncQ repB or IncQ oriTprobes) four IncQ-like plasmids (pIE1107, pIE1115, pIE1120, and pIE1130), each representing a different EcoRV restriction pattern, were selected for a more thorough plasmid characterization after transfer into E. coli K-12 strain DH5α by transformation. The characterization of the IncQ-like plasmids revealed an astonishingly high diversity with regard to phenotypic and genotypic properties. Four different multiple antibiotic resistance patterns were found to be conferred by the IncQ-like plasmids. The plasmids could be mobilized by the RP4 derivative pTH10 into Acinetobactersp., Ralstonia eutropha, Agrobacterium tumefaciens, and P. putida, but they showed diverse patterns of stability under nonselective growth conditions in different host backgrounds. Incompatibility testing and PCR analysis clearly revealed at least two different types of IncQ-like plasmids. PCR amplification of total DNA extracted directly from different manure samples and other environments indicated the prevalence of both types of IncQ plasmids in manure, sewage, and farm soil. These findings suggest that IncQ plasmids play an important role in disseminating antibiotic resistance genes.

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Effect of storage and processing on the immunodetectability of fish proteins using pooled monoclonal antibodies in ELISA and dot blot

Food Control ◽

10.1016/j.foodcont.2021.107976 ◽

2021 ◽

Vol 125 ◽

pp. 107976

Author(s):

Yaozhou Zhu ◽

Yun-Hwa Peggy Hsieh

Keyword(s):

Monoclonal Antibodies ◽

Dot Blot ◽

Fish Proteins

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GLP-1 Analog Liraglutide Improves Vascular Function in Polymicrobial Sepsis by Reduction of Oxidative Stress and Inflammation

Antioxidants ◽

10.3390/antiox10081175 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1175

Author(s):

Johanna Helmstädter ◽

Karin Keppeler ◽

Franziska Aust ◽

Leonie Küster ◽

Katie Frenis ◽

...

Keyword(s):

Oxidative Stress ◽

Vascular Function ◽

Vascular Inflammation ◽

Cecal Ligation ◽

Isometric Tension ◽

Polymicrobial Sepsis ◽

Dot Blot ◽

Markers Of Inflammation ◽

Anti Inflammatory ◽

And Oxidative Stress

Sepsis causes high mortality in the setting of septic shock. LEADER and other trials revealed cardioprotective and anti-inflammatory properties of glucagon-like peptide-1 (GLP-1) analogs like liraglutide (Lira). We previously demonstrated improved survival in lipopolysaccharide (LPS)-induced endotoxemia by inhibition of GLP-1 degradation. Here we investigate the effects of Lira in the polymicrobial sepsis model of cecal ligation and puncture (CLP). C57BL/6J mice were intraperitoneally injected with Lira (200 µg/kg/d; 3 days) and sepsis induced by CLP after one day of GLP-1 analog treatment. Survival and body temperature were monitored. Aortic vascular function (isometric tension recording), protein expression (immunohistochemistry and dot blot) and gene expression (qRT-PCR) were determined. Endothelium-dependent relaxation in the aorta was impaired by CLP and correlated with markers of inflammation (e.g., interleukin 6 and inducible nitric oxide synthase) and oxidative stress (e.g., 3-nitrotyrosine) was higher in septic mice, all of which was almost completely normalized by Lira therapy. We demonstrate that the GLP-1 analog Lira ameliorates sepsis-induced endothelial dysfunction by the reduction of vascular inflammation and oxidative stress. Accordingly, the findings suggest that the antioxidant and anti-inflammatory effects of GLP-1 analogs may be a valuable tool to protect the cardiovascular system from dysbalanced inflammation in polymicrobial sepsis.

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