Hello my fellow peepz!! How have all of you been doing? I’m sure everyone is going through different experiences every single day right? For me, so far, I have learnt more than what I have expected. Even if it is tiring me out almost everyday.

So for this post, I will be talking about what I have been learning in the Clinical Chemistry section.

The basic thing that we need to do is to label and load the samples correctly. Thus, we need to know what tubes is needed for which tests as different tubes is different. For EDTA tubes, it is mainly used for hematology section and GHB. Heparin, Gel and plain tubes can be use for many of the tests. Fluoride tube is used mainly for glucose, particularly fasting.

Before we label, we are to check if the specimens is sufficient to be load onto the machines. If it is more than half of the tube, it is considered sufficient. For the sufficient samples, after labeling, we can load directly into the pre-analytical machine which will be explained in further detail later on.

In this section, there are three main machines or equipments that are used to process the samples. They are the MPA, SWA and Cobas where MPA is used for the pre-analytical processing and SWA and Cobas are used for analysis. Examples of pre-analytical process done on the MPA are centrifugation, aliquoting and labeling. After going through the MPA, the samples will either proceed to the SWA or be aliquot out to be process in the Cobas. So what determines the samples to be run in the SWA or Cobas respectively? Well, it all depends on the different types ordered for that specific samples. Most of the tests can be run on either machines. However, there are some tests that are only specific to SWA or Cobas. Some examples are shown below:

Tests that can only be run in SWA:

- Hepatitis panel

- Cortisol(Serum)

- AFP

- CEA

- PSA

- Pro-BNP

- AHAV total

- HIV

Tests that can only be run in Cobas:

- TDM

- CRP

- GHB

- Anaemia panel 1 & 2

- Magnesium

- Amylase

- Urine tests

- Fluid tests

- C3 & C4

- Direct Bilirubin

- RF

Other tests like renal panel and liver function test can be load on both machines.

There are several circumstances when we are not able to use the MPA for the pre-analytical processing. It could be due to some technical breakdown or when the sample given is insufficient for the MPA to process.

If it is less than half, we are to spin it down and aliquot the serum into the secondary tube. If it is too little, we will use the hitachi cups. Then, we have to load the tube or cups directly to either the SWA or Cobas, depending on the tests ordered. One thing to take note during putting of tubes to each machine manually is that the tubes must not be capped.

For samples requesting for special chemistry tests, the serum is required to be stored in the fridge or freezer. These also depend on the different tests requested. Some examples are shown below:

In freezer – ACTH, PTH and TRAB

In fridge – HbsAg, HCV, HIV, TPO, CA199, Syphillis and Cortisol(urine)

For samples that request for HbsAg, HIV and cortisol(urine), the request form is needed to be kept in the special chemistry bench.

Well, that's all for now.
See you all soon!!
Anw, happy fasting to all of my muslim frens!! Shall meet up to break fast together aite??


Nur Sofieyana
TG02

week 9 of SIP.

hello future med techs. hehe....
hope ur attachment's going fine, mine's okay =)

So now, i would like to share with you a test that is simple but yet important that is done in the Biochemistry Lab.

Test name: G6PD Screening

First, lets just briefly recap on what G6PD is. Glucose-6-phosphate dehydrogenase, G6PD, is a key enzyme needed for the hexose monophosphate pathway which produces NADPH, essential for erythrocyte membrane integrity.This condition might result to destruction of RBCs, leading to hemolytic anemia, followed by jaundice. Thus, for newborns with jaundice, they have their blood sampled to check if the baby's jaundice is caused by G6PD deficiency.

How the test is done:
Patient's sample collected in EDTA tube to prevent clotting.
Patient's tube is overturned 2-3 times to ensure a homogenous composition when sample is taken out.
Using a pipette, 5 microlit of patients whole blood was transferred onto a seperate tube containing 100 microlit of reagent that contains G6-P and NADP.
The mixture is left to sit for 5 minutes.
Below is an equation for the reaction that should take place in the presence of G6PD:

10 microlit of mixture is pipetted out and transferred onto a filter paper.
The filter paper is dried for ten minutes and then viewed under long UV wave in the ulraviolet viewing cabinet.

As seen from the equation above, G6PD is needed for the reaction to take place. The presence of G6PD can be known by viewin the filter paper under the UV light in the ultraviolet viewing cabinet. NADPH flouresces under long wave UV light and a flourescence would indicate the presence of G6PD. In contrast, a sample that does not contain G6PD would not give out any flourescence when viewed under the long wave UV light.



4 controls are set up at the same time:
1. G6PD deficient
2. Intermediate G6PD deficient
3. Bblank (only reagent present)
4. G6PD present



The controls are used as a guide for the patient's results.
For example, a G6PD deficient control does not flouresce at all while an intermediate G6PD control floureses slightly. From the controls, we can gauge the if the patient is completely deficient of G6PD or an intermediate case.



However, this test is just a screen. Samples of patients with positive results are sent out to another laboratory in a hospital for further testing.




feel free to ask ny questions. hope you like my post =)

raihana~

Week Number Eight

Hellohello!!! How are you ppl doing? I hope you guys are doing great. This week, it is my turn again to share my experience. For the past few weeks, I have been rotating about, going to a different lab every week. Quite a lot to absorb for such a short period, but I’ll manage.

I was posted to an infectious disease testing lab one of the weeks. For your information, upon donation, a few samples of blood will be collected from donors to undergo testing, such as infectious disease testing, blood group testing and antibody screening. Under infectious disease itself, it is further divided into a few sections. And the section I am going to explain is TPHA lab.

TPHA is the short form for Treponema pallidum (TP) haemagglutination assay. TP causes Syphilis which is a sexually transmitted disease. The route of transmission is almost always through sexual contact. However, it can also be transmitted through blood transfusions. Therefore, it is important to screen all donors for TP so as to prevent any harmful transmissions and transfusions.

In the TPHA lab, the machine used is the ABBOTT ARCHITECT Syphilis TP. Its intended use is for the diagnosis of Syphilis.

PRINCIPLE OF ASSAY

1. The ABBOTT ARCHITECT Syphilis TP assay is a two-step immunoassay for the qualitative detection of antibody to Treponema Pallidum (TP) in human serum or plasma using Chemiluminescence Microparticle Immunoassay (CMIA) technology with flexible assay protocols, referred to as Chemiflex.
   
2. In the initial step, sample and microparticle coated with recombinant TP antigens (TPN15, TPN17, TPN47) together with the Assay Diluent are combined. Anti-TP antibodies present in the sample bind to the TP coated microparticles. After washing, the acridinium-labelled anti-human IgG and IgM conjugate is added in the second step. Following another wash cycle, Pre-Trigger and Trigger Solutions are added to the reaction mixture.
   
3. The resulting chemiluminescent reaction is measured in relative light units (RLUs). A direct relationship exists between the amount of Anti-TP antibodies in the sample and the RLUs detected by the ABBOTT ARCHITECT i optical system.
   
4. The presence or absence of Anti-TP antibodies in the sample is determined by comparing the chemiluminescent signal in the reaction to the cutoff signal determined from the ABBOTT ARCHITECT Syphilis TP calibration. If the chemiluminescent signal in the sample is greater than or equal to the cutoff signal, the sample is considered reactive for Anti-TP.
   

The specimen used for this assay is either serum or plasma. Haemolysed samples should not be used and all fibrin clots or bubbles must be removed before loading of samples

In the event where there is a positive result, confirmatory tests must be performed. Serum or plasma samples will be collected in a tube and sent to the Serology Laboratory, Department of Pathology, Singapore General Hospital (Reference Laboratory).

The following confirmatory tests are ordered for the determination of the donor’s infection status:

a) Treponema pallidum Particle Agglutination (TPPA) test – an alternative treponemal screening test
b) Venereal Disease Research Laboratory (VDRL) test – a non-treponemal test to determine current or recently treated infection status
c) LIA(Line Immuno Assay) – Syphilis (LIA) – as the confirmatory test for the presence of treponemal antibodies

I hope what I have shared this time round has been a beneficial one. 12 more weeks to go and back to school!! Alright, time to go... see ya

Rusydiana binte Kusni
0608485I
TG02

WEEK 7!!!

Hello everyone!! HOw are your SIP??.. I hope everything's okay, Well I'm having a great in my Micro Lab. For now, I'm in the investigation lab for 1 month where they try to identify the microganisms in the patient's samples. There are quite a number of tests carried out, especially all the biochemical tests( i hope you guys remember). For this week, I learn more about Streptex. I know you guys will be like, "HUH What is Streptex??" . Ok let me tell you...




Streptococcus Latex Agglutination (Streptex)




Streptex is used to identify the Lancefield groups of Streptococci growing on agar plates



Principle


Lancefield showed that the soluble extracted antigens can be identified by precipitation reactions with homologous antisera. Now, latex agglutination or coagglutination methods have largely superseded precipitation method in determining the different groups. A simple enzyme extraction procedure is carried out. Antigen in the resulting extract is identified using polystyrene latex particles which have been coated with group-specific antibodies. These latex particles agglutinate strongly in the presence of homologous antigen and remain in smooth suspension in the absence of homologous antigen.


Materials Streptex Kit

• Streptex Kit contents:Latex for Group A,B,C,D,F,G, extraction enzyme, disposable mixing sticks, disposable reaction cards,


• Disposable Pasteur pipette


• Disposable inoculum loops
  

Procedure

Preparation of extract from culture growing on solid medium:
1. Dispense 0.4ml Extraction Enzyme into an appropriately labelled test tube for each culture to be grouped. (Usually around 20-30 tubes of 0.4ml Extraction Enzyme are prepared at the same time)
2. Using a disposable inoculums loop, make a light suspension of the culture in a tube containing the enzyme solution.( A single sweep of growth or > 5 large colonies will be sufficient to obtain a result)
3. Incubate the suspension at 35⁰C in an incubator for at least 10 minutes


Group Identification:
1. Resuspend each of the latex suspensions by shaking vigorously for about 5 sec (The kit contains 6 bottles, one specific for each of the groups A,B,C,D,F and G)
2. Expel the contents of the in-dwelling droppers to ensure complete mixing.
3. Dispense one drop (20µl) of each latex suspension onto a separate circle on a clean reaction card.
Note: Dropper bottle should be held vertically to ensure that the drops form at the tip of the nozzle. If the nozzle becomes wet, an incorrect volume will be form around the end and not at the tip.
4. Using a Pasteur pipette, place 1 drop of extract in each of the 6 circles on the reaction card.
5. Mix the contents in each circle in turn with a mixing stick, and spread to cover the complete area of the circle. Use a separate stick for each circle and discard it for safe disposal after use.
6. Rock the card gently for a maximum of 1 min, holding it at a normal reading distance (30cm) from the eyes.
7. Discard the card for safe disposal

8. Keep the reagents back into the refrigerator, using the storage rack provided.











Positive result showing agglutination clumps:

Conclusion








Positive result will show development of an agglutinated pattern showing
clearly visible clumping of the latex particles. Speed and quality of agglutination appearance depends on the strength of the antigen extract. Strong antigen extract will give large clumps of latex particles within a few seconds after mixing. Negative result will show no agglutination and the milky appearance remains unchanged throughout the test. There should only be one latex suspensions showing strong rapid agglutination.

So thats about all!! SEE YOU SOON!!!

AmiR ArshaD

TG02

6th week!!!

Olla to all yew gorgeous people! 6 weeks have already gone by and here i am again. Let's cut to the chase shall we?

This week is about....
JENG JENG JENG

SERUM BILIRUBIN~
or at least, testing for bilirubin level in neonatal serum.

As all of us know, babies (especially newborns) have a high probability to be jaundiced. Jaundice during the first 24hrs are usually pathological and are most lkely to be due to either blood group incompatibility or infection. An increase in destruction of RBCs will cause an increase in unconjugated bilirubin, which, in the circulation, will bind to albumin. Once all albumin are fully saturated, the excess unconjugated bilirubin -being lipophillic- can enter the cells. They can also cross the blood brain barrier. There, they can bind to proteins in the brain where it is neurotoxic, and therefore, may result in death or severe mental handicap.

Neonatal jaundice occurs as the liver of the newborns are still incapable of metabolizing the bilirubin. Jaundice in full-term babies usually resolves rapidly. In premature babies, however, the jaundice may be more severe as their liver function is not fully mature.

In the lab I'm attached to, we measure the serum bilirubin using
Wako - Bilirubin Tester II

Analysis Principles:-
Total bilirubin concentration is obtained by measuring the absorbance in the serum. The wavelength is set at 455nm.

Range:-
0 - 1 day 2.0 - 6.0 mg/dL
2 - 5 days 3.9 - 6.0 mg/dL
5 - 127 days 0.3 - 1.7 mg/dL

Specimen:-
2 fully filled heparinized capillary tubes

Equipment:-
Wako-bilirubin Tester II
Centrifuge (3500 rpm/min, 10minutes)

Procedure:-

1. Turn power to ON position & allow 10minutes for lamp to stabilize (Temperature control procedure).
2. Pull out the cell holder and insert a capillary tube (containing distilled water) in front of the slit. Push the cell holder back into the bilirubin tester.
3. Turn the ANA-SET change-over switch to the SET position.
4. Set the Digital Display to ZERO by adjusting the Zero-adjustment control for the 455nm (Filter Selection Lever in the upper position).
5. Using the left hand, push down & hold the Filter Selection Lever in the 575nm (lower position). Set the Digital Display to ZERO by adjusting the zero-adjustment control for 575nm with the right hand.
6. Release the Filter Selection Lever and allow it to return to 455nm position.
7. Pull out the cell holder and set the Digital Display to the standard value by adjusting the Span-adjustment control.
8. Turn the AVA-SET chenge-over switch to the ANA position.
9. Insert a capillary tube filled with sample (already centrifuged) into the cell holder.

10. Push down & hold the Filter Selection Lever into the 575nm posititon. Confirm that the Digital Display at ZERO, then allow level to return back to 455nm position.
11. Read the Digital Display & read results.
12. To measure subsequent samples, repeat steps 9 - 11.

All pictures taken with permission from supervisor.

So, there you go. Seems complicated, but very simple really. Any doubts, feel free to ask =)

Enjoy your SIPs people!!!!

Name: Mayafirhana

Class: TG02

LMQA

Name: Mayafirhana Bte Hairulhassan
Subject title: Laboratory Management and Quality Assurance
Topic: Organization or regulatory bodies that regulate the operations of local laboratories in Singapore

The Workplace Safety and Health (WSH) Council collaborates with the Ministry of Manpower (MOM) to help other industries to achieve 'A safe and healthy workplace for everyone; and a country reowned for best practices in workplace safety and health'.1

The WSH Council:

- Help recognize and practice proper safety and health in the employees when working.2

- Created the Construction Safety Audit Scoring System (ConSASS) which offers an independent evaluation of the safety and health management system at the company.3

- Provides surveillance to keep track and control the health of the employees, and the hazards found at the worksite.4

1. Workplace Safety and Health Strategy 2015. (2008). Retrieved 31 July, 2008, from: http://www.wshc.gov.sg/WSH2015.html


2. The Expansion of the WSH Act. (2008). Retrieved 31 July, 2008, from: http://www.wshc.gov.sg/pub_guidelines.html


3. About ConSASS. (2008). Retrieved 31 July, 2008, from: http://www.wshc.gov.sg/ConSASS.html


4. Health and Environmental Surveillance. (2007). Retrieved 31 July, 2008, from: http://mom.gov.sg/publish/momportal/en/communities/workplace_safety_and_health/maintaining_a_safe_workplace/health_and_environmental.html

Laboratory Management and Quality Assurance

Name: Mohamed Amir bin Mohamed Arshad

Subject Title: Laboratory Management and Quality Assurance

Topic: Organization or regulatory bodies that regulate the operations of local laboratories in Singapore

Ministry of Health (MOH) regulates clinical laboratory in Singapore, providing consistent, safe and affordable healthcare services(1) to all Singaporeans by implying to various Legislative Acts.

The regulating acts ranges from healthcare professional to related medicinal substances.(2)
There is the Medicines Act which consists of suitable administration, quality and analysis of drugs, and good clinical practices. (3)

MOH implements new regulatory framework to make sure Singaporeans are updated on healthcare services. For example, in the recent liposuction cases, MOH maintains the safety and confidentiality of the patients. Hence any clinic that carries out liposuction must acquire approval from the Ministry. (4)

1. About MOH. (2007). Retrieved 31 July, 2008, from: http://www.moh.gov.sg/mohcorp/about.aspx?id=82

2.Legislation. (2007). Retrieved 31 July, 2008, from: http://www.moh.gov.sg/mohcorp/legislations.aspx?id=214

3.Medicine Acts (2008). Retrieved 31 July 2008, from:
http://statutes.agc.gov.sg/non_version/cgi-bin/cgi_retrieve.pl?actno=REVED-176&doctitle=MEDICINES%20ACT%0A&date=latest&method=part

4.Liposuction Regulatory Framework (2008). Retrieved 31 July 2008, from:
http://www.moh.gov.sg/mohcorp/default.aspx