Sandwich nucleic acid hybridization: A method with a universally usable labeled probe for various specific tests

Wolf, Hans; Leser, Ulrike; Haus, M.; Gu, S. Y.; Pathmanathan, R

doi:10.1016/0166-0934(86)90066-2

Cited by 14 publications

(5 citation statements)

References 8 publications

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“…Single stranded D N A could be directly biotinylated (i.e. by photobiotinylation) [5,, indirectly detected by a second biotinylated probe [19], or used as template to obtain single stranded probes by primer extension [12-1, allowing even higher levels of detection. These alternatives are currently being tested.…”

Section: Discussionmentioning

confidence: 99%

Biotinylated nucleic acid hybridization probes for potato virus detection

Hopp

Giavedoni

Mandel

et al. 1988

Archives of Virology

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cDNA libraries, representative of potato viruses X (PVXc strain) and Y (PVY degrees strain) genomes were obtained. A PVX cDNA cloned fragment was sequenced and biotinylated to be used as hybridization probe for the detection of purified virus or nucleic acid extracts of infected plants. Dot hybridization assay was sensitive to detect 4 ng of viral particles, corresponding to about 200 pg of viral RNA. The level of detection in infected plant extracts was as effective as that obtained with the ELISA. The presence of biotinylated PVY cDNA in the hybridization mixture did not affect sensitivity of the PVX detection assay, suggesting that a single diagnostic assay for several potato viruses and virus-related pathogens could be developed.

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Section: Discussionmentioning

confidence: 99%

Biotinylated nucleic acid hybridization probes for potato virus detection

Hopp

Giavedoni

Mandel

et al. 1988

Archives of Virology

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“…Such a scheme using Sephacryl S-500 has been used as a rapid method to distinguish between DNA from sickle, heterozygote and normal patients [33]. Very recently it has also been applied successfully to typing human papilloma, viruses with the considerable advantages of speed, low cost and potential automation over the more conventional procedure of dot blotting, hybridization with a A further sandwich hybridization (indirect) has been suggested recently by Wolf et al [65] and applied to the detection of cytomegalovirus in urine specimens. It makes use of M13 probes.…”

Section: Nucleic Acid Hybridizationmentioning

confidence: 96%

“…Viral inserts are placed in the intergenie M13 sequence which does not affect viral replication and inserts up to 1000 nucleotides are stable on replication of the bacteriophage. One example of an 3/[13 recombinant probe being used diagnostically is for Rhinovirus detection in nasal secretions [2], and an indirect sandwich hybridization based on M13 probes is described below [65].…”

Section: Preparation Of Nucleic Acid Probesmentioning

confidence: 99%

Advances in the use of nucleic acid probes in diagnosis of viral diseases of man

Norval

Bingham

1987

Archives of Virology

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A variety of methods are now available for the preparation and labelling of viral nucleic acids for use as probes in diagnostic virology. Some of these are assessed including the use of synthetic oligonucleotides in place of molecularly cloned nucleic acids, and alternatives to labelling with radioactive isotopes such as biotin, enzymes and fluorochromes. Dot blot, sandwich, indirect sandwich and in situ hybridization are covered, and examples given of the current use of nucleic acid probes in detection of human viral infections. The potential and limitations of nucleic acid hybridization are discussed in the light of these new methods.

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“…As the spectrum of nucleic-acid-based assays expands, so the number of different probes required will increase, re-sulting in logistical problems of manufacture and storage; this will be particularly marked where isotopes with short half-lives are used. This difficulty may be surmounted by using a sandwich hybridization in conjunction with a universally applicable probe (94). An unlabelled M 13 construct containing a 3,000-bp insert specific for cytomegalovirus (CMV) AD169 DNA was hybridized to a sample immobilized onto nitrocellulose (stage 1).…”

Section: Variations On the Sandwich Themementioning

confidence: 99%

Nucleic acid analysis by sandwich hybridization

Nicholls

Malcolm

1989

Clinical Laboratory Analysis

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P e t e r J o h n N i c h o l l s . A T h e s i s S u b m it t e d f o r t h e D e g r e e o f D o c t o r o f P h i lo s o p h y o f t h e U n i v e r s i t y o f L o n d o n . C h a r i n g C r o s s a n d W e s t m in s t e r M e d i c a l S c h o o l.M a r c h , 1 9 8 9 . 2 A b s t r a c t . T h e n u c l e i c a c i d -b a s e d t e s t s c u r r e n t l y u s e d f o r t h e d e t e c t io n o f i n h e r i t e d a n d i n f e c t i o u s d i s e a s e a r e s e n s i t i v e a n d s p e c i f i c , b u t n o t a lw a y s r e p r o d u c ib le , b e c a u s e o f p r o c e d u r a l c o m p l e x i t i e s ; t h i s i n t u r n m eans t h a t t h e y a r e n o t a m e n a b le t o a u t o m a t io n . S u c h t e s t s a r e n o t c o s t -e f f e c t i v e , a r e l i m i t e d i n t h e num ber o f s a m p le s w h ic h c a n b e p r o c e s s e d a t o n c e , a n d m o s t com m only r e l y o n u n s t a b le i s o t o p i c a l l y l a b e l l e d p r o b e s , w h ic h a r e h a z a r d o u s t o h a n d le a n d d i s p o s e o f . A n u c l e i c a c i d s a n d w ic h h y b r i d i z a t i o n a s s a y h a s b e e n d e v e lo p e d , i n i t i a l l y u s i n g c lo n e d hum an p a p i l l o m a v i r u s (H PV) s e q u e n c e s a s a m o d e l, a n d s u b s e q u e n t ly f o r t h e d e t e c t io n a n d t y p i n g o f H PV i n f e c t i o n i n c e r v i c a l s c r a p e s . T h e i m p l i c a t i o n s o f c e r v i c a l HPV i n f e c t i o n a r e o f c l i n i c a l s i g n i f i c a n c e , b e c a u s e o f t h e r e l a t i v e l y r e c e n t a c c u m u la t io n o f e v id e n c e l i n k i n g s p e c i f i c H PV t y p e s w i t h t h e a e t i o l o g y o f c e r v i c a l c a n c e r . C o n s i d e r i n g t h a t v i r a l t y p e c o u ld h a v e a g r e a t i n f l u e n c e o n t h e p r o g n o s i s o f i n f e c t i o n , a n d t h a t im m u n o lo g ic a l a s s a y s a r e n o t c a p a b leo f d i f f e r e n t i a t i n g b e tw e e n t y p e s , t h e s a n d w ic h h y b r i d i z a t i o n t e c h n iq u e , w h ic h c o u ld h a n d le l a r g e n um bers o f s a m p le s , w o u ld b e a n i n v a l u a b l e t o o l f o r im p r o v in g t h e e f f i c a c y o f t h e c e r v i c a l s c r e e n i n g p rogram m e. T o Mun a n d D ad, w ith a l l n y lo v e . T h e a s s a y r e q u i r e s tw o d i s t i n c t , n o n -o v e r la p p i n g ENA fr a g m e n t s , d e r iv e d fr o m a d j a c e n t p o s i t i o n s o n t h e v i r a l genom e. One f r a g m e n t (A ) i s c o v a l e n t l y a t t a c h e d t o a S-T h a n k s f o r m a k in g t h i s p o s s i b l e - 5T h e kncw n i s f i n i t e , t h e unknow n i n f i n i t e ; i n t e l l e c t u a l l y we s t a n d o n a n i s l e t i n t h e m id s t o f a n i l l i m i t a b l e o c e a n o f i n e x p l i c a b i l i t y . O u r b u s i n e s s i n e v e r y g e n e r a t io n i s t o r e c l a i m a l i t t l e m ore la n d .T .H . H u x le y , 1 8 8 7 . F i r s t a n d f o r e m o s t ,...

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Sandwich nucleic acid hybridization: A method with a universally usable labeled probe for various specific tests

Cited by 14 publications

References 8 publications

Biotinylated nucleic acid hybridization probes for potato virus detection

Biotinylated nucleic acid hybridization probes for potato virus detection

Advances in the use of nucleic acid probes in diagnosis of viral diseases of man

Nucleic acid analysis by sandwich hybridization

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