Microbiology Laboratory Turkey

Mikrobiyoloji Ile Ilgili Tüm Konuların Kısa ve Öz Anlatımları. Microbiology Lab Information.

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MicroLab

29 Mayıs 2019 Çarşamba

Mayıs 29, 2019

Dientamoeba fragilis

Dientamoeba fragilis

Dientamoeba fragilis was initially classified as an amoeba; however, the internal structures of the trophoziote are typical of a flagellate. No cyst stage has been described. The life cycle and mode of transmission of D. fragilis are not known. It has worldwide distribution. The transmission is postulated, via helminthes egg such as those of Ascaris and Enterobius species. Transmission by faecal- oral routes does occur. Most infection with D. fragilis is asymptomatic, with colonization of the cecum and upper colon. However, some patients may develop symptomatic disease, consisting of abdominal discomfort, flatulence, intermittent diarrhea, anorexia, and weight loss. 
The therapeutic agent of choice for this infection is iodoquinol, with tetracycline and parmomycine as acceptable alternatives. The reservoir for this flagellate and  lifecycle are unknown. Thus, specific recommendation for prevention is difficult. However, infection can be avoided by maintenance of adequate sanitary conditions. 

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15 Mart 2019 Cuma

Mart 15, 2019

Kızamık Virüsü | Morbillivirus measles virus | RUBEOLA | Rubella | Kızamıkçık

Kızamık Virüsü | Morbillivirus measles virus | RUBEOLA | Rubella | Kızamıkçık

120-150 nm boyutlarında, tek iplikli, zarflı, RNA virüsüdür.

Antijenik Yapısı:
Bir tek antijenik tipi vardır. Kızamık antijeni enfekte hücreler içinde tespit edilmiştir. Antijenleri kompleman birleşmesi deneyi ile de tespit edilebilir
Kızamık Virüsü:
Kızamık hastalığının etkenidir. Hastalık bir kez geçirilmekle ömür boyu bağışıklık bırakır. Virüse karşı canlı atenüe aşı kullanılır. Kızamık, kızamıkçık, kabakulak olarak üçlü aşı (MMR) şeklinde uygulanır.  


Teşhis Yöntemleri:
Genellikle klinik belirtilerle kolayca tanı konur.

Seroloji:
Klinik belirtilerle teşhis edilemeyen vakalar için en çok kompleman birleşmesi deneyi kullanılır. Ayrıca fluoresan antikor ve ELISA testleri de yapılır.

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14 Mart 2019 Perşembe

Mart 14, 2019

Oxidation-Fermentation (OF) Test (or Hugh-Leifson Test)

Oxidation-Fermentation (OF) Test (or Hugh-Leifson Test)

The Oxidation-Fermentation (OF) Test tests the metabolism of sugar by prokaryotic cells.  Cells may metabolize sugar in a variety of pathways, and the OF Test is studying whether sugar is metabolized by aerobic respiration or by an anaerobic pathway including fermentation.  The OF medium has a low agar and peptone content, and a high sugar content, making it a semi-solid medium that is unlikely to go alkaline from protein utilization if the sugars are metabolized.  Sugar fermentation produces fairly strong acids while aerobic respiration produces only weak acids, either of which will turn the indicator, Bromthymol Blue, yellow.  Bromthymol Blue is green around a neutral pH and blue at an alkaline pH (>7.5).  

Two tubes are stab inoculated with an organism, then one tube is covered with mineral oil, creating an anaerobic environment to study fermentation.  The other tube is left uncovered with oil, creating an aerobic environment to study aerobic respiration.  Organisms that do not utilize the sugar in both tubes will leave the media blue or green (non-saccharolytic, non-sugar user).  Organisms that only engage in aerobic respiration will turn the media yellow in the top of the uncovered (aerobic) tube while the covered tube remains blue or green.  

Organisms that only slowly ferment the sugars will be slightly yellow at the top of both tubes.  Organisms that slowly ferment the sugars and aerobically respire the sugars will also be slightly yellow at the top of both tubes.  Organisms that strongly ferment the sugars will be yellow throughout both tubes.  Organisms that both stronly ferment and aerobically respire the sugars will also be yellow throughout both tubes.  Obviously, this test can only differentiate organisms into three or four groups:

  1. Non-sugar metabolisers
  2. Aerobically respiring organisms
  3. Organisms that either aerobically respire and ferment or ferment only
  4. And organisms that either aerobically respire and slowly ferment or slow fermenters only
➤The semi-solid medium also allows for the detection of motile organisms which will radiate out from the stab.

Aim of Test: To differentiate bacteria species on their ability of aerobically respire or ferment sugar and to determine motility.  This test allows for the differentiation of the fermenting Enterobacteriaceae from the aerobically respiring Pseudomonas and Bordetella and from the non-sugar using Alcaligenes and Moraxella

Procedure:
  1. Obtain two tubes for each strain being tested (your instructor will inform you of which strains to test).  
  2. Label the tubes, including your name, the date, and the test name.  
  3. Do NOT use your Wire loop, as this will soon ruin it. Use your inoculating needle for this test.
  4. If assaying for motility, care must be taken with the stab.  Be certain that your needle is straight and that your stab is straight down and then pulled up exactly the way it went down.
  5. Aseptically transfer some bacteria to the inoculating needle.  This is done by flaming your needle, COOLING IT WELL, picking up bacteria on the needle by rolling the tip in a colony of bacteria (oe scaping it).  Sometimes students get no growth in this test from just briefly cooling the tip of the needle: the heat from above the tip moves down the needle and kills the bacteria.  So coool it well.
  6. Stab the needle straight down into the tube going about 2/3 of the way down and then pulling the needle straight out the way you came in.
  7. A poor stab will make the motility reading difficult, so repeat the test if needed.
  8. Repeat this procedure for the second tube of the set.
  9. Overlay one tube of each set with a thin layer of sterile mineral oil (about 4 mm).
  10. Remind your instructor to set up a control tube (an uninoculated control).  Gently place your tubes in the rack where instructed.  Rough handling may distort the test.
  11. Incubate the tubes at room temperature for about 5 days (or at 35-37 C for 2 days).
Results:
  1. Examine the tubes for color changes, noting where the color has changed.  If tubes have no visible growth and have not changed color or the organism is slow growing, additional incubation time may be required.
  2. Look for motility as in the motility test.
  3. Check your results against the table below.
Note to the Instructor: 
Although at first glance this test looks pretty interesting, it actually adds little but confirmation to the knowledge obtained from the Utilization of Glucose Test and the Thioglycollate Test. If given a choice between the two, the latter tests are recommended instead of the OF Test.

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13 Mart 2019 Çarşamba

Mart 13, 2019

Anti-HCV Testi | Kaset Testi

Anti-HCV Testi

Hepatit C virüsü flaviviridae ailesinden olup,  zarflı ve tek sarmallı RNA içeren bir virüstür. Hepatit B gibi parenteral yoldan bulaşır. Ağırlıklı olarak virüs taşıyan kan ve kan ürünlerinin kişiden kişiye nakli yoluyla geçmektedir. Diğer bulaşma şekilleri ise damar yoluyla uyuşturucu kullanımı, cinsel ilişki ve enfekte anneden hamilelikte bebeğe virüsün geçmesidir. 
Günümüzde Anti-HCV testi mikrobiyoloji laboratuvarında otoanalizörde ve ticari olarak değişik firmaların ürettiği “kaset test”lerle bakılmaktadır. 
Hepatit C virüsüne karşı oluşan antikorları 14-30 gün arasında tespit edilir. Bu test immünokromatografik prensibe dayanmaktadır. 
Amaç
İnsan serum ya da plazmasında Hepatit C virüsüne karşı bağışıklık sistemi tarafından üretilen antikorların kalitatif olarak tespit edilmesidir.  

Araç Gereçler
 Viyalda (ısıya dayanıklı plastik malzemeden yapılmış çoğunlukla vida kapaklı numune saklama tüpü) 
 Santrifüj 
 Anti HCV test kaseti 
 Tek kullanımlık plastik damlalık 
 Taze insan serum ya da plazması 

Teknik
 Test kaseti oda sıcaklığına getirilir. 
 Test başlamadan hemen önce test kaseti poşetinden çıkarılıp, düz bir zemin üzerine yerleştirilir. 
 Numune deliğine 2-3 damla serum / plazma damlatılır. 
 Kasetlerin numuneyi emmesi beklenir. 

Testin Değerlendirilmesi
 Pozitif sonucu en geç 10 dakika sonra okuyunuz. 
 Negatif sonucu en geç 20 dakika sonra okuyunuz. 
 20 dakikadan sonra alınan sonuçlar dikkate alınmamalıdır. 
 Sonuç penceresinde sadece “C” harfi üzerinde kalan bölümde yer alan tek çizgi negatif sonucu gösterir. Bu, numunenin HCV virüsüne karşı antikor barındırmadığını gösterir. 
 Sonuç penceresinde hem “C” hem “T” harfleri üzerinde kalan bölümde yer alan çizgiler pozitif sonucu gösterir. Bu, numunenin HCV virüsüne karşı antikor içerdiğini gösterir. 
 Sonuç penceresinde beklenmesi gereken süre sonunda hiçbir çizgi görünmez ise sonuç geçersizdir. Test prosedürü hatalı uygulanmış ya da test arızalanmış olabilir. Yeni bir test ile deney tekrarlanmalıdır

Anti-HCV Kaset Testinde Dikkat Edilmesi Gereken Kurallar
 Kullanmadan önce kullanma talimatını mutlaka okuyunuz. 
 Testleri kesinlikle dondurmayınız. Eğer test buzdolabında saklanmış ise ürünü kullanmadan önce oda sıcaklığına getiriniz. 
 Deney oda sıcaklığında (15-30 oC) yapılmalıdır. 
 Son kullanma tarihi geçmiş testleri kullanmayınız. 
 Test kasetlerini poşetinden çıkartınca hemen kullanınız. 
 Test kasetlerini poşeti yırtılmış ise kullanmayınız. 
 Deney esnasında tek kullanımlık eldiven kullanınız 
 Materyalin ağız, yüz, göz ve açıkta bulunan yara ile temas etmemesine özen gösteriniz. 
 Deney esnasında yiyecek tüketmeyiniz ve sigara içmeyiniz. 
 Hemolitik, lipemik veya bakteri içeren numuneleri kullanmayınız. 
 Test kasetlerinin verdiği sonuçlar mutlaka ek tanı ve teşhis yöntemleriyle uzmanlar tarafından desteklenmelidir.

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12 Mart 2019 Salı

Mart 12, 2019

Hydrogen Sulfide (H2S) Production Test

Hydrogen Sulfide (H2S) Production Test

This test can be performed with the use of several media including Triple Sugar Iron (TSI), Kligler's Iron Agar (KIA), and SIM medium. 

 An iron compound and a sulfur compound are included to test for the production of hydrogen sulfide gas.  Hydrogen sulfide is produced if the sulfur compound is reduced by the bacterial strain.  This happens when the strain either degrades the amino acid cysteine during protein degradation, or when anaerobic respiration shuttles the electrons to sulfur instead of to oxygen.  In either case H2S is produced (hydrogen sulfide gas) which reacts with the iron compound to form the black precipitate of ferric sulfide.  The black color acts as an indicator for the presence of hydrogen sulfide.  (If the tube becomes largely black, it may be difficult to read the tube for other tests.)


Purpose:
This test aids in the identification and differentiation of members of Enterobacteriaceae (enterics) from other Gram- bacilli.  It is especially helpful in identifying Salmonella, Francisella, and Proteus species.


Procedure:
  1. Refer to the medium that you are using for the procedure as they differ by medium.
  2. Observe the medium in the tube and record its color and appearance in your notebook.
Results:
  1. Look for the occurrence of H2S production (black coloration), comparing your tube to an uninoculated control if needed.
  2. The black coloration may mask the color of the the tube, especially the lower end.
  3. Make your interpretations and record your results and interpretations.

Instructor Notes:
TSI is less sensitive in detecting the production of hydrogen sulfide gas than other similar tests (SIM media, Kligler's Iron Agar, etc.).  So, it is possible to observe the production of hydrogen sulfide in another test and not observe it in TSI.  The sucrose in TSI may suppress the formation of hydrogen sulfide.  Both TSI and Kligler's Iron Agar also use ferrous sulfate to detect the production of H2S, and this detector is less sensitive than other hydrogen sulfide detectors.  TSI has an added sulfur source (sodium thiosulfate), but sulfur may also be obtained from certain amino acids present in the protein in the media.

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9 Mart 2019 Cumartesi

Mart 09, 2019

HYMENOLEPIS DIMINUTA (RAT TAPEWORM)

HYMENOLEPIS DIMINUTA
(RAT TAPEWORM)

Hymenolepis diminuta differs from Hymenolepis nana in that: 
♦ The adult worm measures about 10-60 cm 
♦ The rosetellum on the head has no hooks 
♦ In the mature segment, there are two testes at one side and another testis on the other side.
Life Cycle
The adult worms are present in the small intestine of man and rats. Eggs passed in stool are similar to the eggs of H. nana but are brown in color with no polar filaments arising from the polar thickening. The eggs are ingested by the rat flea where they develop to cysticercoid stage. Infection to man takes place accidentally by food or contaminated hands by cysticercoid stage.
Pathogenecity
Most infections are asymptomatic, but occasionally, patients may present with nausea, anorexia and diarrhea.

Treatment
Same as Hymenolepis nana.

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