Detection of Human Rotaviruses in Fresh and Estuarine Waters by Dot-blot Hybridization

1991 ◽  
Vol 23 (1-3) ◽  
pp. 253-260 ◽  
Author(s):  
Abidelfatah M. Nasser ◽  
Mary K. Estes ◽  
Theodore G. Metcalf
Keyword(s):  
Fresh Water ◽  
Water Samples ◽  
Hepatitis A Virus ◽  
Blot Hybridization ◽  
Genome Segment ◽  
Detection Sensitivity ◽  
Dot Blot ◽  
Cross Hybridization ◽  
Dot Blot Hybridization ◽  
Stool Samples

This study was conducted to evaluate the suitability of dot-blot hydridization for detecting rotavirus in fresh and estuarine water samples using cloned DNA of genome segment 6 of human strain Wa and simian SA-11 rotaviruses. Probes of Wa and SA-11 cross-hybridized with heterologous RNAs of SA-11 and Wa rotaviruses under low stringency conditions. However, cross-hybridization reactions were minimized or eliminated under high stringency conditions, while homologous reactions were not affected. Wa probe did not react with representatives of the major enterovirus groups or hepatitis A virus. Hybridization detection sensitivity was not affected by organic material present in waters or proteinaceous material of eluents. Hybridization using cDNA of Wa rotavirus was found to be more sensitive than either Rotazyme ELISA or electron microscopy for detecting rotavirus in stool specimens. Dot-blot hybridization using cDNA of genome segment 6 of Wa rotavirus was found to be more sensitive than SPRIA for detecting rotavirus in estuarine and fresh water samples. Only 9 out of 54 (18.5%) estuarine and fresh water samples were positive by SPRIA, whereas dot-blot hybridization with Wa probe resulted in 21 out of 54 (38.8%) positive samples. One out of 20 (5%) stool samples and 23 out of 54 (44.6%) estuarine and fresh water samples were found to be positive by a pBR-322 plasmid probe. These results indicate that pBR-322 and related plasmids are present in high frequency in fecally polluted waters. Although different numbers of positive results were found with Wa and pBR-322 probes, more research is required to ascertain that hybridization reactions with such plasmids do not occur when using cDNA rotavirus probes.

Characterization of Marine Temperate Phage-Host Systems Isolated from Mamala Bay, Oahu, Hawaii

1998 ◽  
Vol 64 (2) ◽  
pp. 535-542 ◽  
Author(s):  
Sunny C. Jiang ◽  
Christina A. Kellogg ◽  
John H. Paul
Keyword(s):  
Water Samples ◽  
Blot Hybridization ◽  
Temperate Phage ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Double Stranded Dna ◽  
The Family ◽  
Marine Viruses

ABSTRACT To understand the ecological and genetic role of viruses in the marine environment, it is critical to know the infectivity of viruses and the types of interactions that occur between marine viruses and their hosts. We isolated four marine phages from turbid plaques by using four indigenous bacterial hosts obtained from concentrated water samples from Mamala Bay, Oahu, Hawaii. Two of the rod-shaped bacterial hosts were identified as Sphingomonas paucimobilis andFlavobacterium sp. All of the phage isolates were tailed phages and contained double-stranded DNA. Two of the phage isolates had morphologies typical of the family Siphoviridae, while the other two belonged to the families Myoviridae andPodoviridae. The head diameters of these viruses ranged from 47 to 70.7 nm, and the tail lengths ranged from 12 to 146 nm. The burst sizes ranged from 7.8 to 240 phage/bacterial cell, and the genome sizes, as determined by restriction digestion, ranged from 36 to 112 kb. The members of the Siphoviridae, T-φHSIC, and T-φD0, and the member of the Myoviridae, T-φD1B, were found to form lysogenic associations with their bacterial hosts, which were isolated from the same water samples. Hybridization of phage T-φHSIC probe with lysogenic host genomic DNA was observed in dot blot hybridization experiments, indicating that prophage T-φHSIC was integrated within the host genome. These phage-host systems are available for use in studies of marine lysogeny and transduction.

scholarly journals Species-Specific Detection of the Maize Pathogens Sporisorium reiliana and Ustilago maydis by Dot Blot Hybridization and PCR-Based Assays

Plant Disease ◽  
1999 ◽  
Vol 83 (4) ◽  
pp. 390-395 ◽  
Author(s):  
M. L. Xu ◽  
A. E. Melchinger ◽  
T. Lübberstedt
Keyword(s):  
Genomic Dna ◽  
Blot Hybridization ◽  
Pathogenic Fungi ◽  
Ustilago Maydis ◽  
Dot Blot ◽  
Cross Hybridization ◽  
Dot Blot Hybridization ◽  
Fungal Dna ◽  
Species Specific ◽  
Sporisorium Reiliana

Head smut of maize, caused by Sporisorium reiliana, may substantially reduce grain yield. The objective of the present study was to develop a highly specific and sensitive DNA-based assay for detection of S. reiliana and its differentiation from Ustilago maydis, a maize fungus inducing the symptomatically similar common smut disease. Plasmid libraries of S. reiliana and U. maydis were constructed using a shotgun cloning procedure. Clones containing strongly hybridizing species-specific DNA were selected by screening libraries with their own labeled genomic DNA, followed by cross-hybridization with genomic DNA of maize and other maize-pathogenic fungi. The selected clones were used to generate subclones with short insert fragments to facilitate PCR amplification for labeling and primer design for a PCR assay. Using Dig-dUTP labeled inserts, detection of less than 0.16 ng of fungal DNA was possible by dot blot hybridization. Sequences of insert fragments were determined to design primer pairs for a PCR-based assay. Primer pairs SR1 and SR3 are species-specific for S. reiliana, and UM11 is species-specific for U. maydis. The PCR-based assays can detect fungal DNA of less than 1.6 pg using SR1 and SR3, and 8 pg using UM11, irrespective of the presence of maize DNA. Use of SR1 and SR3 allowed detection of S. reiliana in the extracts of pith, node, and shank from S. reiliana-infected plants, but not in leaves. Thus, both the dot blot hybridization and the PCR-based assays provide a highly sensitive and reliable tool for detection and differentiation of corn smut caused either by S. reiliana or by U. maydis.

scholarly journals A dot-blot hybridization procedure for the detection of astrovirus in stool samples

1991 ◽  
Vol 107 (2) ◽  
pp. 405-410 ◽  
Author(s):  
M. M. Willcocks ◽  
M. J. Carter ◽  
J. G. Silcock ◽  
C. R. Madeley
Keyword(s):  
Electron Microscopy ◽  
Nucleic Acid ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Hybridization Procedure ◽  
Hybridization Test ◽  
Stool Samples

SUMMARYWe have developed a nucleic acid dot-blot hybridization test for the detection of astroviruses in stool samples. The test was not as sensitive as electron microscopy for the detection of low numbers of well preserved astrovirus particles, but was able to identify astroviruses in stools containing particles of indistinct morphology. In total, this procedure identified astroviruses in more samples than did electron microscopy, and the data indicate that the incidence of astroviruses may be substantially underestimated.

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

1991 ◽  
Vol 24 (2) ◽  
pp. 267-272 ◽  
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.

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)

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

Prevalence of HPV in a Melbourne Female STD Population: Comparison of RNA and DNA Probes in Detecting HPV by Dot Blot Hybridization

1993 ◽  
Vol 4 (3) ◽  
pp. 159-164 ◽  
Author(s):  
A J Borg ◽  
G Medley ◽  
S M Garland
Keyword(s):  
Past History ◽  
Blot Hybridization ◽  
Condylomata Acuminata ◽  
Dot Blot ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Dot Blot Hybridization ◽  
History Of ◽  
Cytological Features ◽  
Dna Types

A total of 377 women, consecutively selected as first attenders to a sexually transmitted diseases clinic in Melbourne, Australia, were examined for overt Condylomata acuminata and were screened for genital HPV DNA types 6, 11, 16, 18, 31, 33 and (35) using 2 dot blot hybridization methods. Overall, there was a 90% positivity correlation between the 2 methods with HPV DNA being detected in 12% of ectocervical samples. Overt warts were found in 15% of the women and HPV DNA was detected at the cervix in 35% with cytology predicting HPV with or without dysplasia in 27%. Thirteen percent had a past history of warts but none on examination and HPV DNA was evident in 16% while 18% had cytological features of HPV. Those with no warts evident and no past history of warts had both HPV DNA and cytological features of HPV in 7%.

Detection of herpes simplex virus and adenovirus DNA by dot blot hybridization using in vitro synthesized RNA transcripts

1986 ◽  
Vol 13 (4) ◽  
pp. 291-299 ◽  
Author(s):  
Volker Schuster ◽  
Bertfried Matz ◽  
Helga Wiegand ◽  
Brigitte Traub ◽  
Dieter Neumann-Haefelin
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Rna Transcripts ◽  
Dot Blot Hybridization ◽  
Simplex Virus

Detection of Cryptosporidium parvum Oocysts on Fresh Vegetables and Herbs Using Antibodies Specific for a Cryptosporidium parvum Viral Antigen

2005 ◽  
Vol 68 (5) ◽  
pp. 1093-1096 ◽  
Author(s):  
K. E. KNIEL ◽  
M. C. JENKINS
Keyword(s):  
Cryptosporidium Parvum ◽  
Viral Antigen ◽  
Blot Hybridization ◽  
Surface Antigens ◽  
Food Samples ◽  
Sensitive Detection ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Fresh Vegetables ◽  
Cryptosporidium Parvum Oocysts

The purpose of this study was to determine if the viral symbiont of Cryptosporidium parvum (CPV) sporozoites could be used as a target for sensitive detection of the parasite in food samples. Polyclonal sera specific to the recombinant viral capsid protein (rCPV40) was used in a dot blot hybridization assay to detect oocysts recovered from green onions and cilantro. Small batches of chopped green onions and cilantro leaves were artificially contaminated with three different concentrations of oocysts: 106, 102, and 101. rCPV40 was superior in detecting oocysts compared with other antibodies directed toward total oocyst protein and oocyst surface antigens. This study provides evidence that CPV is an excellent target for sensitive detection of C. parvum oocysts in foods.

Sign in / Sign up

Export Citation Format

Share Document