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1
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• ?????? 500 ? 300 ?????? ??? ?? ?? ????? ?? ????
  (80 ???? ?? ??????? ????????)
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  ????? ?? ?? ?? ??? ?? ???? (80 ???? ??? ??? ??? 5
  ???).

2

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  ???? ??? ?????? ?????.

3
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4
(No Transcript)
5

• ?????????? ?????????? ???????? ????? ???? ?? ???
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• ?????????? ?????? ? ?????????? ?????? ?????? ????
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6

• ?????????? ?????? ???? ???? ???? ?????????? ??
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7

• ??? ?????? ?????????? ?????? ? ??????? ???????
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  ????? .
• ????? ???? ? ???? ?????????? ??? ?????.

8

• ?? ???????? ???? ?? ?????????? ??????? ?? ?? ????
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  ??? ?? ??? ????? ???? ?????.
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  ?????????? ?????? ?? ?? 4? 3 ??? ?? ??? ?? ????

9

• ?????????? ?????? ?? ???? ???? ???? ?? ???? ??
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  ????? ?? ???.

10

• ???? ? ?????????? ?? ??? 17? 10 ??? ??? ?? ???.
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• ???? ????? ???????? ???? ?? ?????? ??? ?? ?????
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• ??? ?? ?????? ???? ?????? ???????? ????? ????????
  ??? ?? ???? ??? ???? ???? ???? ?? ????? ???.

11

• ???? ? ????? ? ?????? ?? ?????? ?? 30 ? 9 ???
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• ???? ?? ?????? ?????????? ?????? ?? ???? ?????
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• ??? ???? ???? ?? ?????????? ?????? ? ???? 10 ??
  17 ???? ?? ?????????? ??????? 18 ?? 40 ??? ? ??
  ?????????? ?????????? 11 ? 8 ??? ???.

12

• ?????? ?? ?????? ????? ????? ??? ????? ????? ???
  ????? ?????? ???? ????? ? ????? ?????? ????
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13

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  ?????????? ??????? 72 ???? ???.

14

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  ??????? ????? ? ??????? ?? ????? 5/5? 8? 9 ? 15
  ???
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  ??????????? 12 ? 8 ??? ?? ?????? ? 35 ??? ??
  ??????? ?? ????.

15

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16

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17

• ???????? ???? (????? ????? ????? ???? ???????
  ????) ? ?????? ???????? (?????? ??? ?????? ??? ?
  ?????? ?????? ??? ??????? ??? ??? ? ????).

18

• ?? ???? ???? ?
• ??????? ???? (???? ???? ?? ??????)?
• ??? ???? (???? ???? ???? ?? ????? ?????)?
• ???? ??? ??? ?
• ???????? ??? ( ???? ???? ???? ???? ???? ????
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• (??????? ???? ?? ??? ??? ???? ? ?????? ?????)?
• ?? ????? ???? (?????? ?? ??? ?????? ???
  ????????? ?? ?????) ?
• ????? ????.

19
?????

• ????? ???? ??????? ?? ??????? ?????????? ???? ??
  ????? ???? ???? ?? ????.
• ???????? ???? ??? ????? ???? ??? ?? ??? ????? ??
  ??? ???? ????? ????? ??? ???????? ??????????
  ?????????? ?? 10 ??? ?? ?? ???? ????? ??? ??????
  ???? ??? ???.

20

• ????? ???? ??? ????? ???? 15 ????? ???? ?????
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  ????? ?? ???? ???? ????? ?? ?? ???? ????? ???.
• ????? ???? ???????? ??? ?????? ???? ??????? ???.

21
Types of Serological Assays Malaria

• Antibody Detection
• Indirect Fluorescent Antibody
• Enzyme immunoassays
• Antigen Detection
• Immunochromatographic

22
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23
Indirect Fluorescent Antibody (IFA)
24
Quantitative Buffy Coat (QBC )

• Useful for screening large numbers of samples
• Quick, saves time
• In the QBC technique, approximately, 50
  microlitres of whole blood was taken into a
  capillary tube coated with acridine orange and
  fitted with a cap.

25

• A plastic float was inserted inside the tube and
  then spun in the QBC microhaematocrit centrifuge
  at 12000 rpm for 5 minutes
• Due to acridine orange, the malarial parasite
  stains green (DNA nucleus) and orange (RNA
  cytoplasm).

26

• The tube is examined in the region between the
  red blood cells and granulocytes and within the
  granulocytes and mononuclear cell layer, where
  parasites are most abundant.

27

• main disadvantages
• 1-Species identification and
• 2-quantification difficult
• 3-Requires centrifuge, special stains

28
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29
Malaria Antigen Detection

• Immunologic assays to detect specific antigens
• Commercial kits now available as
• immunochromatographic rapid diagnostic tests
  (RDTs), used with blood
• P. falciparum histidine-rich protein 2 (PfHRP-2)
• parasite LDH (pLDH)

30

• Monoclonal and polyclonal antibodies used in
  antigen (Ag) capture test
• Species- and pan-specific Ab
• Cannot detect mixed infections
• Cross reactivity with rheumatoid factor
  reportedly corrected

31
Detection of Plasmodium antigens pLDH (parasite
lactate dehydrogenase)
32
Detection of Plasmodium antigens
A HRP-2 (histidine-rich protein 2) B pLDH
(parasite lactate dehydrogenase C HRP-2
(histidine-rich protein 2)
33
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34
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35
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36
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37
Polymerase Chain Reaction (PCR)

• DHFR Gene (dihydrofolate reductase-thymidylate
  synthase )
• Molecular technique to identify parasite genetic
  material
• Uses whole blood collected in anticoagulated tube
  (200 µl) or directly onto filter paper (5 µl)

38

• 100 DNA is extracted
• 10 blood volume used in PCR reaction

39
Polymerase Chain Reaction (PCR)

• Threshold of detection
• 0.1 parasite/µl if whole blood in tube
• 2 parasites/µl if using filter paper
• Definitive species-specific diagnosis now
  possible
• Can identify mutations try to correlate to drug
  resistance
• Parasitemia not quantifiable
• May have use in epidemiologic studies
• Requires specialized equipment, reagents, and
  training

40
Sensitivity of Tools forDiagnosis of Malarial
Infection

• Most sensitive
• Antibody detection
• PCR
• Blood film examination

41
Diagnosis of Untreated Acute Malaria

• Blood film examination
• PCR

42
Diagnosis of Treated Recent Malaria

• Serology
• Blood film examination
• PCR

43
Diagnosis of Chronic Malaria

• Screen with serology
• If IFA positive
• May do blood film examination
• May do PCR

44
Mass Screening for Malariain Populations for
Resettlement

• Blood smear examinations to detect asymptomatic
  parasitemia
• Not useful for predicting individual risks
• undetectable parasitemias
• dormant liver phase parasites
• Can be used to make a decision about the need for
  mass treatment of the entire group

45
Other tests

• Rapid reduction in level of hemoglobin is seen in
  falciparum malaria and less than 7 g/ dl should
  be a warning.
• Leukocytosis is vary from low to high, and
  neutrophilic leukocytosis is common in severe
  malaria with or without associated bacterial
  infection.

46

• Thrombocytopenia is common in P. falciparum and
  P. vivax malaria
• Hypoglycemia is a common problem encountered in
  malaria
• Hyperbilirubinemia is common in malaria,
  particularly due to hemolysis
• Serum albumin levels may be reduced, some times
  markedly.

47

• Hyponatremia is common and needs careful
  management.
• Prothrombin time and partial thromboplastin time
  are elevated in 20 of patients with cerebral
  malaria

48
?????

• ????? ????? ???? ???? ???????? ???? ?? ???? ???
  ??? ?????????? ?????????? ? ??? ???????? ???? ??
  ????? ???? ?? ??? ???? ?? ?? ?? ??? ????? ?????
  ???? ?????? ??????? ????? ?????? ??? ?? ?? ?????
  4? ???????????? ?? ????.

49

• ?? ????????? ???? ???? ?? ???? ??? ???? ?? ?????
  ?????????? ?????????? ? ?? ?? ????? ???????
  ??????? ?????? ???? ??? ????? ??????? ????????
  ????? ???? ????.
• ???? ??????? ?? ?????? ???? ?? ?????????? ??????
  ? ?????? ?? ????? ???? ?? ?? ???????? ????? ?? ??
  8 ? ???????????? ?? ???? ???? ???
• ??? ???? ?????????? ??? ???? ?????????? ?????? ?
  ???? ?? ?? ??? ???? ? ???? ??? ?????? ?? ?? ????.

50

• ???? ????? ?????????? ?????????? ????? ?? ???????
  ???? ????? ?????? ????? ???? ???? ???.
• ?? ????? ?????? ?? ??????? 3 ??? ???????? ??
  ???? ?? ?? ????? ?? ???.
• ??? ???? ?????? ?? ????????? ? ??????????? ??
  ????.
• ?? ???? ????? ???? ???? ?? ???????? ?? ?????
  ????? ?????? ?? ?????????? ?? ????? ? ?? ????
  ???? ????? ?? ??????? ???? ?? ????? ????? ???????
  ?? ???.
• ?????? ???????? ??????? ? ??????? ??????? ??????
  ?? ???? ? ???????? ?? ?????.

51
?????? ??????

• RI ???? ???? ??? ??????? ?? ??? ??? ????? ? ????
  ?? ??? ? ??? ?????? ?? ?? ??? ?? ??? ?? ?? ????
  ????? ???? ?? ???.
• RII ?????? ???? ????? ???? ?? ??? ?? ?? ????
  ????? ??? ??? ??? ??? ??? ? ?????? ???? ?????
  ???? ???.

52

• ?? RIII? ???????? ??? ?????? ? ?? ?? ????? ?????
  ?????? ?? ??? ?? ???? ????? ???? ??? ??? ????
  ?????? ?? ???? ?? ???.
• ?? ?????? ??? RI ?? ???? ??????? ?? ????? ?????
  ???.
• ?? ???? ???? ?????? ?? ???? ?? ??????? ????
  ?????? ??? ???????? ??????? ???.

53
??? ??? ?????

• ???? ????? ???? ???? ?????? ??? 9? 2 ???? ? ?????
  ???? ???? ???.
• ???? ????? ?? ????? ????? ?? ???? ????? 9? 2 ????
  ?? ??? ?? ?? ?? ????? ???? ??? ???? ??????????
  ???? ???.
• ?????? ?? ???? ????? ????? ?? ?? ?? ????? ?????
  ??????? ???? ????? ????? ??????? ???? ??
  ????????? ???? ?? ????.

54

• ?? ????? 2 ????? ???? ??????? ???? ????
• ????? ???? ???? ?????? ????? ? ????? ???? ??.
• ?? ????? ???? ?? ?? ?????? ?????? ??? 1950
  ??????? ????? ???? ? ?????? ???? ???? ???? ???
  ???? ? ?? ????? ?? ???? ????? ?? ??? ?????? ????
  ?????? ?? ????.

55

• ???? ????? ??????? ?? ??? ????? ????? ?? ????????
  ?? ???? ?? ?? ???? ???? ?? ??? ??? ???? ??? ???.
• ????? ??? ????? ?? ????? ???? ??? ??? ?? ? ???
  ?????? ?? ???? ?? ?? ? ??? ???? ??? ???? ???????
  ? ??????? ?? ??? ???? ????? ??????? ?? ???? ?????
  ?????.

56

• ?? ????? ???? ????? ??????? ?? ? ?????? ??????
  ??? ?? ?? ???? ?? ??? ?????? ???? ??? ?????? ?
  ?????? ????( ??? ?? 12? 10) ???? ? ?????? ??????
  ? ??? ??????? ? ??????? ? ??????? ????? ??? ???
  ??? ??? ?? ??????? ?? ??????? ???? ??? ???? ????
  ????.

57

• ?? ??? ???? ???? ?? ???????? ?? ??? ?????? ????
  ??????? ????? ??? ??? ??? ?? ????? ???? ???? ????
  ????? ?????? ? ????????? ??? ????? ??????? ? ????
  ????? ????? (????? ? ?????) ???? ???? ???????
  ???? ????.

58

• ?? ????? 15 ???? ????? ?? ??? ????? ???? ???? ??
  7 ???? ???? ???? ?? ?????? ?????? ??????? ??
  ?????????
• ????? ??????? ??? ???? ???? ?? ????? ????? ?????
  ?? ????.
• ????? ????? ?????? ??????? ??????? ?????? ???? ?
  ??? 50 ?? 100 ???? ???.
• ?? ???? ???? ????? ?? ???? ????? ????? ?? ?? ???
  ?????????? (???? ?? 10 ????)? ????????? (11 ?? 50
  ????) ? ??????????75-50 ?? ?? ???? ???? ???.

59
???????

• ??????? ???? ????
• ??? ???? ??????? ? ???? ????? ???????? ??? ??
  ??? ????? ????? ?? ?? ??? ? ??? ????? ????????.
• ?????? ?????????
• ??? ??? ??? ???? ??? ? ??????? ???? ?? ??????? ?
  ??? ???????? ??? ????? ?? ??? ?? ??? ??????
  ??????? ?? ???.

60

• ?????? ?????? ?????
• ???? ??????? ?? ???? ???????? ?? ???? ?? ???????
  ?? ?? ????? ????? ??? ????? ??? ???? ?????? ??
  ??? ?? ???? ???? ?? ??? ??????? ??? ???? ?? ???.
• ???? ?????? ?? ??? ??????? ???????? ??? ????? ??
  ??? ?? ??? ??? ??????? ?? ???.
• ?????? ?? ??? ????
• ?? ???? ?? ?? ??? ???? ?? ????.

61

• ?? ???? ?????? ????? ??????
• ?? ?? ???? 2 ?? 6 ??? ?? ? ???? ???? ?? ???? ?
  ?? ????? ????? ?? ???? ????? ? ?????? ??????? ??
  ???.
• ?? ???? ????? ????? ???? ????.
• ?? ????? ????? ???????? ???? ?? ?????? ?? ????
  ???? ???? DDT(?? ???? ?? ???? ??? ???? ????)?
  ???????? ? ??????? ?????? ???? ???? ???.

62

• ?????? ??????? ?? ??????? ???? ??? ???????? ?? ??
  ????? ???? ?? ???? ??? ?? ?????? ???? ?? ???? ???
  ??? ??????? ????? ???? ????.
• ??????? ?? ??? ???
• ??????? ?????? ?? ??? ??? ?? ???? ?????? ?? ??
  ?? ????????? ????? ???? ?? ??????? ???????? ?????
  ???? ???.

63
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64
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65
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66
(No Transcript)
67
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68
(No Transcript)
69
(No Transcript)
70
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71
(No Transcript)
72
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73
(No Transcript)