Infeksi Gastrointestinal non diare
Dian Widiyanti, SSi., MSi, Ph.DBagian Mikrobiologi
Fakultas Kedokteran Universitas YARSI
Helicobacter pylori
HistoryBarry Marshal & Robyn
Warren in 1982 Campylobacter pyloridis from dyspepsia patient now known as Helicobacter pylori
In 2005 nobel prize in physiology or medicine
MorphologySpiral with 1-3 turns, 0.5 ×5 μm in
length, with 5 to 7 polar sheathed flagella
Motile with corkscrew motilitySlow growth (3-6 days) at 37o CpH optimum 6-7Microaerophilic organismOxidase positive, catalase positive,
produce ureaseCause acute or chronic gastritis,
predisposing factor in peptic ulcer, gastric carcinoma, MALT lymphoma
Transmission
Risk factor
Virulence Factor Effect
Colonizing
Flagella Active movement through mucin
Urease Neutralization acid
Adhesin Anchoring to epithelium
Tissue damaging
Proteolytic enzymes Glucosulfatase degrades mucin
120-kDa cytotoxin (Gac A) Related to ulcer and severe gastritis
Vacuolating cytotoxin (Vac A) Damage of the epithelium
Urease Toxic effect on epithelial cell, disrupting tight junction
Phospholipase A Digest phospholipid in cell membrane
Alcohol dehydrogenase Gastric mucosal injury
Survival
Intracellular surveillance Prevent killing in phagocytes
Superoxide dismutase, catalase Prevent phagocytosis and killing
Coccoid forms Dormant form
Heat shock protein
Urease Sheathing antigen
Other
Lipopolysaccharide Low biological activity
Lewis X/Y blood group homology Autoimmunity
Pathogenesis
Clinical sign and symptoms
Upper abdominal painNausea and vomitingFrequent burpingBloatingLoss appetite, Fast satiationWeight lossBleeding from stomach lining (severe case)
Stages in Helicobacter infection
H. pylori induce autoimmune
Diagnosis Non invasive test
Urea breath test Stool antigen Serology (ELISA)
• Endoscopic based test
Rapid urease test Histology Culture (Br+5%
horse blood or BHI+ 7% horse blood) 5-10% O2, 5-12% CO2 5-7 days at 37o C
Treatment
Kombinasi beberapa antibiotik + inhibtor pompa proton
Contoh amoksilin, metronidazol, omeprazol
Leptopirosis
Occupational hazard of rice harvesting (China), autumn fever (Japan)
Adolf Weil in 1886 the first modern clinical description of leptospirosis which characterized by splenomegaly, jaundice, nephritis Weil’s disease
Inada et.al in 1916 • isolated leptospires, • identified the organism as the causal agent of leptospirosis • determined that rats are reservoir for transmission to human
Classification Order: SpirochaetalesFamily : Leptospiraceae
L. interrogans sensu lato (>240 serovar, 24 serogroups)L. biflexa sensu lato (>60 serovars)
Genotypic classification 20 genomospecies
Morphology• Spiral-shaped bacterium• Size 0.1 x 6-20 m• Highly motile• Has two axial filament (endoflagella) at its ends• Obligate aerobes • Slow growth with optimal temperature 30o C• Has Gram-negative bacterial cell wall
Reference: Albert I. Ko, et.al., 2009, Nature Reviews Microbiology, 7:736-747
(Reference: Jurg Utzinger, et.al., 2012, Swiss Medical Weekly 142:w13727)
Reservoir host
Cattle Dog Sheep and Goat
Horses Pig
Leptospira serovars
PomonaHardjo
Canicola PomonaHardjo
BratislavaPomona
BratislavaAustralisPomona
Symptoms and syndromes
Reproductive failureAbortionStill-birthsFetal mummificationWeak calvesMilk drop syndrome
IctericHemorrhagicUremic (Stuttgart disease)Abortion and premature or weak pups
SepticemiaRedwaterAbortionMilk drop syndrome
AbortionRedwater in foalsMoon blindness
AbortionStill-birthWeak piglets
Leptospirosis in Animals
Mice (Mus musculus) and rats (Rattus norvegicus and Rattus rattus) don’t show sign but harbor leptospires in kidney Hamsters and guinea pigs are highly susceptible and can be used as animals models of human leptospirosis
Leptospirosis in humans
Reference: Feigin RD, Anderson DC: Human leptospirosis. CRC Crit Rev Clin Lab Sci 1975;5: 413-67. Copyright CRC Press, Inc., Boca Raton, FL
PathogenesisThe clinical manifestations are caused by damage to the
endothelial lining of small blood vessels by mechanisms that are still poorly understood. Infection multisystem. Some virulence factors hemolysin, Lig A and Lig B, LPS
TreatmentPenicillin i.v., amoxicillin, ampicillin, doxycycline, eritromicinJarisch-Herxheimer reaction may appear following penicillin treatment
Prevention Prevention and control should be targeted at :
(a) the infection source; (b) the route of transmission between the infection
source and the human host(c) infection or disease in the human host.
(Reference: Pappas G., P. Papadimitriou, V. Siozopoulou, L. Christou, and N. Akriditis. 2008. Int. J. Infect. Dis. 12: 351-357
The Caribbean and Latin America, the Indian subcontinent, Southeast Asia, Oceania, and to a lesser extent Eastern Europe, are the most significant foci of the disease
Clinical sample for leptospirosis diagnosis
1. BloodBlood is used for culture, collected with heparin to
prevent clotting on the first 10 days. Ten days after the onset, leptospires mostly dissappeared from blood.
2. SerumSerum is used for serologic test, collected two
times at interval several days. To detect rise in titres between two samples or seroconversion and confirm the diagnosis
3. UrineMidstream urine is collected and cultured as soon as possible (2 hours after sampling) because leptospires
die quickly in urine.4. Postmortem
Collected from many organs soon after the death and transported to laboratorium in 4o C to prevent autolysis.
5. CSF
Diagnosis of leptospirosis
Detection of host responseMicroscopic Agglutination Test (MAT)
one of the gold standard in diagnosis of leptospirosis. Serum of suspected patient is reacted to culture of leptospira strain. The agglutinated of leptospires will form clumps. MAT result is
determined positive if the proportion of free leptospires is less than 50%. Enzyme-Linked Immunosorbent Assay (ELISA)
using wide variety of leptospiral sonicates to recombinant protein (i.e. LipL32, LigA, OmpL1). IgM dipstick
detect the IgM which appeared during acute phase and often use for initial screening.
Microcapsule Agglutination Test (MCAT)
Detection of causative agentCulture
growth in EMJH or Korthof medium. Definitive diagnosis, but need long incubation due to slow growth of Leptospira strains
Polymerase Chain Reaction (PCR) has high sensitivity
Loop-mediated isothermal amplification (LAMP) amplification of DNA under isothermal condition and the result can be seen with naked eye. It has high sensitivity and specificity, no need sophisticated equipment.
Latex agglutination, immunofluorescence, immunoprecipitation lack of sensitivity and specificity
WHO References1. The culture of sample from normally sterile organ, showed
sufficient growth2. Clear amplified DNA in PCR from organ or body fluid3. Four-fold increase of titers between acute and convalescent sera in
MAT