Dengue Virus Plaque Formation in Rhesus Monkey Kidney Cultures

Georgiades, J; Stim, Thomas B.; McCollum, Robert W.; Henderson, J. R.

doi:10.3181/00379727-118-29851

Cited by 9 publications

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“…Since the introduction of cell culture systems, many people have developed various assay systems for DEN viruses. The principle of these methods is either by plaque assay (Schulze and Schlesinger, 1963;Miles and Austin, 1963;Georgiades et al, 1965;Hotta et al, 1966;Westaway, 1966;Russell et al, 1967;Bergold and Mazzali, 1968;Banerjee, 1969;Yunker and Cory, 1975) with its adaptation to micromethod (Skuhavachana et al, 1969;DeMadrid and Porterfield, 1969;Fujita et al, 1975) or to hemadsorption negative plaque (Makino and Mifune, 1975), or by interference method (Halstead et al, 1964). We have developed the focus counting method for the rapid titration of DEN viruses (Igarashi and Mantani, 1974;Okuno et al, 1977Okuno et al, , 1978.…”

Section: Discussionmentioning

confidence: 99%

Comparative Sensitivity of Focus Counting Method in the Infectivity Titration of Dengue Viruses and in the Neutralization Test

Igarashi

Okuno

Fukai

1979

Jap. J. Trop. Med. Hyg.

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Abstract:Infectivity titrations were performed for each of the 4 prototypes of dengue (DEN) viruses by focus counting on BHK21 cells (FFU), by plaque formation on LLC-MK2 cells (PFU), and by intracerebral inoculation into suckling mice (LD50) The FFU titers were almost similar as LD50 for all the 4 types of DEN and these titers were also similar to PFU except type 2 DEN (DEN-2), which showed lower PFU than FFU or LD50. Neutralization (N) tests were performed for standard anti-DEN sera by 50 per cent focus reduction (FR50) and by 50 per cent plaque reduction (PR50) methods. Both values were almost the same except anti-DEN-2, whcih showed higher PR50 than FR50. The principle of these methods were also applied to Japanese encephalitis and chikungunya viruses. Hemagglutination-inhibition (HI) and FR50-N tests on several human sera indicated that FR50 is more specific and sensitive than HI to detect typespecific antibodies.

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Section: Discussionmentioning

confidence: 99%

Comparative Sensitivity of Focus Counting Method in the Infectivity Titration of Dengue Viruses and in the Neutralization Test

Igarashi

Okuno

Fukai

1979

Jap. J. Trop. Med. Hyg.

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“…Virus assay. Procedures of virus inoculation and preparation of nutrient agar overlay were reported previously (4,12). Virus dilutions for titrations were made in Hanks saline containing 0.5% lactalbumin hydrolysate.…”

Section: Materiais and Methodsmentioning

confidence: 99%

“…d Plaques developed but not clear enough to count. DISCUSSION Although dengue virus plaquing was first accomplished by using methyl cellulose (2%) as the solidifying agent in the overlay (9), agar of various purities and concentrations (0.7 to 2.0%) has been preferred by most laboratories for dengue plaquing (4,12,13). Unpurified agar (Difco) was not acceptable for optimal dengue plaquing, nor Expressed as logio plaque-forming units per 0.2 ml.…”

Section: 6mentioning

confidence: 99%

“…The concentrations used were based on toxicity level experiments carried out with LLC-MK2 and Vero cells. Specific plaque development data of all dengue serotypes, in the absence of plaque-potentiating compounds for the LLC-MK2 cell system, were presented in a previous paper (4).…”

mentioning

confidence: 99%

“…Effect of using old molten agar and diethylaminoethyl (DEAE) on dengue type4 in LLC-MK2 cellsValues after agarb kept at 56 C for days"…”

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confidence: 99%

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Dengue Virus Plaque Development in Simian Cell Systems

Stim

1970

Applied Microbiology

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Dengue type 2 virus, strain New Guinea B, plaqued with equal facility and titer under overlays containing six different grades of commercial agar in the LLC-MK2 cell system. Doubling the agar volume on LLC-MK2 cell monolayers increased the plaque development time of dengue type 1, strain Hawaii. Storage of agar at 56 C reduced or totally abolished dengue type 4, strain H-241, plaque titer in LLC-MK2 cells. The influence of six known virus plaque-enhancing compounds on plaque development of all dengue virus serotypes was studied in two continuous simian kidney cell lines, LLC-MK2 and Vero. In the absence of any chemical additive, plaque development of all dengue serotypes was more rapid (4 to 10 days) in the LLC-MK2 line than in the Vero line (6 to 13 days). Increased plaque development time of type 1, strain Hawaii, by pancreatin and plaque-size doubling of dengue types 1 and 4 was the only advantage conferred by the addition of six chemical additives in the LLC-MK2 cell system. Dengue types 1 and 6 failed to plaque in the Vero cell system unless aided by a plaque-enhancing compound; plaques of dengue types 2, 3, 4, and 5 appeared sooner (2 days) and were increased in plaque diameter. The optimal DEAE concentration for plaquing dengue type 1, strain Hawaii, was 100 ,g/ml; plaque development either failed at lower concentrations or was inhibited at higher (200 ,g/ml) concentrations.

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Effects of Invading Organisms on Cells and Tissues in Culture

Bang

1966

Cells and Tissues in Culture Methods, Biology and Physiology

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Dengue Virus Plaque Formation in Rhesus Monkey Kidney Cultures

Cited by 9 publications

References 0 publications

Comparative Sensitivity of Focus Counting Method in the Infectivity Titration of Dengue Viruses and in the Neutralization Test

Comparative Sensitivity of Focus Counting Method in the Infectivity Titration of Dengue Viruses and in the Neutralization Test

Dengue Virus Plaque Development in Simian Cell Systems

Effects of Invading Organisms on Cells and Tissues in Culture

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