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biotechnology > speciation > Flashcards

speciation Flashcards

1
Q

what is speciation

A

establishment of species by origin by the use of analytical techniques

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2
Q

how is speciation done

A

sampling of tissue, food, fluid or other organic matter

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3
Q

what is the aim of speiciation

A

determine from a sample what species the sample came from

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4
Q

why is speciation important

A

food fraud
medical testing
forensics
protection of rare spieces

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5
Q

what types of approaches are used for speciation

A

utilise genetic differences between species via PCR or RT PCR - based on the gene

immunologic assays - ELISA, biosensors - Based on the gene product

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6
Q

food production is a —— pound industry

A

multi billion

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7
Q

what type of legislation is in place to protect the consumer

A

extensive

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8
Q

when were food laws passed about food adulteration

A

recently

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9
Q

how can we now detect and monitor food fraud

A

recent technological advancments

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10
Q

what is adulterated foods

A

food containing poisons, colouring, other additives to disguise, inferior quality or where an important constituent has been substituted

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11
Q

what is the most common for of adulteration

A

substitution of cheaper ingredient for a more expensive one

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12
Q

when was food adulteration a main problem

A

19th century

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13
Q

what was tea normally padded out with

A

gun powder

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14
Q

what was the most adulterated staple

A

bread - add boiled potato, sawdust. chalk, clay, bone dust,

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15
Q

cream thickened with what

A

calf brains

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16
Q

name the methods of speciation

A

antibody based
DNA based
Infer-red spectroscopy
mass spectroscopy

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17
Q

why use an ELISA for speciation

A 
widely used in food 
compared to other methods it is highly sensitive
cheap
rapid
requires little training
antibodies readily available
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18
Q

what is an ELISA

A

quantitative method used to measure antigens and antibodies
based on antigen/antibody interactions
antibodies produced in immune resoponce
antibodies are specific to a particular antigen

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19
Q

what is an antibody

A

glycoprotein that recognise and bind to a particular antigen with very high specifity

20
Q

advantages of polyclonal antibodies

A

broad specify

cheap

21
Q

disadvantages of polyclonal

A

finate supply

22
Q

advantages of monoclonal antibodies

A

narrow specifity

continuous supply

23
Q

disadvantages of monoclonal antibodies

A

expensive

24
Q

advantages of recombinant antibodies

A

narrow specify

genetic manipulation

25
Q

disadvantages of recombinant antibodies

A

expensive

poor stability

26
Q

what is a recombinant antibody

A

bacteria
fungi
plants

27
Q

what is a monoclonal antibody

A

mouse
rat
hybridoma cells

28
Q

what is a polyclonal antibody

A 
rabbit
sheep
goat
house
chicken
29
Q

advantages of ELISA

A 
qualitative and quantitative
detect antigen and how much is present 
can detect in serum, urine, saliva etc
detect antibodies to indicate disease (HIV)
can also use for environmental testing
30
Q

what will a specific assay only detect

A

desired analyse - won’t give positive results with any other protein

31
Q

what is the limit of detection

A

lowest amount of antigen needed to be detected

32
Q

the lower the detection limit the —-

A

more sensitive the elisa

33
Q

describe reactants in an ELISA

A

once bound stay attached to the solid surface.

washing steps- resulting in the removal of unbound reagents allow easy measurement of bound complex.

34
Q

what are ELISA normally carried out in

A

96 well micro titre plate allowing large analysis of large number samples

35
Q

what is the normal means of measurement in an ELISA

A

enzyme labelled component

normally peroxidase or phosphate conjugates

36
Q

simple ELISA is —-

A

direct detection

37
Q

Indirect ELISA is —

A

indirect detection

38
Q

sandwich ELISA is —-

A

direct detection

39
Q

competitive ELISA is

A

both direct and indirect

40
Q

what are the disadvantages of an simple ELISA

A

primary antibody, must be labelled with enzyme or other label

uses a lot of antibody, unsatisfactory when primary antibody is in short supply/ expensive

41
Q

describe advantages of simple ELISA

A

quick
simple
few steps
avoids potential cross reactivity which may occur when using secondary antibodies

42
Q

advantages of indirect ELISA

A

labelled secondary antibody
increases signal strength as several labelled secondary antibodies can bind to primary

a single second antibody can be used with variety of primary antibodies made in same species

secondary antibody is species specific

43
Q

disadvantages of indirect ELISA

A

more steps than direct

potential cross reactivity of secondary antibody

44
Q

describe direct sandwich ELISA

A

antibody imobilised into plate to capture antigen and extract from a sample

antigen can be directly detected using labelled primary antibody or indirectly using primary antibody followed by labelled secondary antibody

45
Q

advantages of sandwich ELISA

A

highly specific
used to extract minute amounts of antigen which is too small to be directly absorbed onto micro titre surface
highly sensitive

46
Q

Disadvantages of sandwich ELISA

A

require 2 antigen specific antibodies

many steps involved

47
Q

common problems associated with ELISA include

A

non specific binding - can be due to other substance in sample, limited by blocking plate

interference binding - due to other substance in sample

problems with reagents

  • antibodies
  • many buffers used

biotechnology (6 decks)

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