Sputum Contamination and Centrifugation

This is the very first step in Culture specimen for diagnosis and identification Tuberculosis. Sputum decontamination is the process of removing contamination, particularly the cleaning off of dangerous materials (sputum). Sputum specimen was collected in falcon 50 ml in 2 collection times ie morning an in place/health provider (labeled with P and S). In this study, we examine sputum smear negatives sent by 3 BP4 (Balai Pengobatan Penyakit Paru-Paru) in Yogyakarta.
The decontamination is processed in the Basic Safety Containment level 2 (BSC 2) to protect the laboratory workers from the infection risks, since the transmission of TB can be through aerosol. The lab workers should wear the protection equipments: lab jacket, gloves and mask. The steps are described below:
1. Mix NaOH 3% and Na Citrat with the same volume (25+25 ml)
2. Add NALC 0.25 gr (after mix with NaOH-Na Citrat should be used within 24 hours)
this is known as decontamination solution.
3. Add the solution into specimen with volume 1:1
4. mix in vortex mixer
5. Add PBS (Phosphate Buffer Saline) until volume is 45 ml
6. centrifuge in 4 degree, 3500 RPM for 15 minutes.
7. Throw the all of the supernatant and add with PBS 2ml
8. Mix with vortex mixer
The sputum has been decontaminated and centrifuged and ready to be inoculated in the medium. In this study the decontaminated specimen is inoculated in Thin Layer Agar (with and without Para Nitro Benzene/PNB), Lowenstein Jensen (with and without PNB) Mycobacteria Growth Indicator Tube (MGIT) and object glass for direct microscopic examination. The rest of the specimen is kept in the pellet and freeze in -20 degree to be used for other purposes examination.

Firs day in Laboratory

After a week I've been busy with settling everything in Jogja, finally I can breath freely and start my journey. I visited Medical Faculty GMU to meet Bu Ning, my supervisor who has given me this opportunity. Nine o'clock in the morning sharply I stepped in to laboratory's door. Unfortunately, she has'nt come yet. I tried to enter the door after waiting for 15 minutes outside. I knocked one of the doors and meet Ibu Sunyi. She was smiling and trying to recognize me. She still remembered me and in a short time we had a long chat. She is the only one assistant of Bu Ning who works for TB research. She gave me some references and explained to me the progress of the on going research that I will involve in.
Let me give you an overview about the research. This research is about " Thin Layer Agar as a new rapid diagnosis of TB in smear negative patients in Jogjakarta", conducted by Department of Tropical Medicine GMU in collaboration with Institute of Tropical Medicine, Antwerp, Belgium. Since I will attend a TB course in Lausanne in October, they ask me to visit Antwerp for gaining some knowledge and skills in Thin Layer Agar. This is a good opportunity for me to start a new journey in TB Lab. I will assist Bu Ning in lab working for the on going study. The objective of the study is determine the sensitivity and specificity of TLA compare with other methods (Lwenstein Jensen/LJ and Mycobacterium Growth Indicator Tube/MGIT) in smear negative patients. The target samples is around 1400 negative smears patients taken from 3 BLKs in Jogjakarta. 161 has been successfully collected and examined.
After waiting for 3 hours, finally Bu Ning came in to her room. She looked happy and of course busy. She told me about the research and asked to observe the lab works for this 2 weeks and do it by myself after. Having half an hour effective discussion with her made me conclude that I should work hard with this.
Fortunately soon after the discussion was ended, I had an opportunity to observe directly the preparation of media (TLA), sputum decontamination and inoculation process in to TLA, MGIT, LJ and object glass for BTA examination. I will explain latter on the steps of each method. Basically, I really enjoy it and excited to involve in this study.
As I always say to Lita: I will never stop learning. Thanks for opening my way God, I will try more harder.

Smart Risk

I watched a kid's program accidentally in abckids this morning. His program has made me realize what happen in my life in this short time. Many value things that I need to share here.

The host asked the audience: do you feel bored if everyday you do the same things in your life? eat, sleep, watch, eat, sleep and watch again. We called this as a comfort zone. If you want to do something new, we called it take a risk. It is not always as dangerous risk but a smart risk. Sometime get out of the comfort zone make us feel uncomfortable. Therefore, before you decide to take the smart risk you should assess the positive and negative things that will be affected to the risk. From this assessment you should determine what is your goal and what are you afraid of? How you can cope this feeling and take the first step?

This is really useful for me to be applied in my recent new decision. I am assuming it as my smart risk. My goal is to get a new experience in TB area and enhance my capability and knowledge. The positive thing that might be happen if I take this risk is I can start my new carrier that is related to my background. The negative thing that might be happen is they don't satisfy with the result of the course. Thus, that is one thing that I am afraid of: failure.

For reducing the negative thing to be happened, I should more reading, learning and keep practicing as much as I can. Do my best and let God taking care of the rest.

Biosafety

1. Definition

Biosafety consist of 2 words: Bio (biology) and safety, means protection to potentially harms from infection agents including.

Infective Microorganism were classified into 4 risk group:

Risk group 1 (no/low individual and community risk)

Risk group 2 (moderate individual and low community risk)

Risk Group 3 (high individual and low community risk)

Risk Group 4 (high individual and community risk)

If there is a limitation of information, the epidemiological data and patient history can be used to asses the risk.

Biosafety level is divided into 4 level based on the composite of design features, construction, containment facilities, equipment, practices and operational procedures.

Biosafety level1 - basic

Biosafety level 2 - basic

Biosafety level 3 - containment

Biosafety level 4 - maximum containment

The backbone of Biosafety is Risk Assessment that performed by professional judgement as the laboratory director responsibility. This should be reviewed routinely and revised if required.

All Izz Well

Komang Andika, my brother in law, has arrived in Wollongong yesterday. Andi picked him up in the Sydney airport by train. We know that winter season is not a good time for holiday for him because of its extremity in weather difference with Bali. In the morning the weather can be very cold below 5 degree even sometimes minus 2 degree then 10 to 15 degree in the mid day. In contrast to Bali, the weather mostly hot that vary from 30-40 degree. Hope he can adapt soon with this kind of situation so he can enjoy his journey here. Imagine how excited Lita when she knew his "Om Upik" will come. She looks happy, talkative and asks more attention from Upik and ayah. She show how she can eat by herself and explain everything she knew to Upik.

We watched an Indian movie brought by Upik last night. "3 Idiots", a quite long duration movie like other Indian movies but we really enjoy it. It was a story about friendship between 3 students in a college for mechanical engineering, Rancho, Raju and Farhan. They are come from a different family background. Raju's dad used to be a postman but his stroke make him laying on the bed, meanwhile mum working hard for taking care of her family. His family hope he can be engineer so can take offer the family backbone and earn money for his sister marriage. Unfortunately his marks always at the bottom ranks in the class, and his friends called him idiot. Meanwhile Farhan, is a good son from middle class family, but his interest is in wild animals photography instead of engineering. His father think that he can not have money from that kind of profession and push him to be an engineer. Raju's marks in the class is slightly different with Farhan, ranks number 2 from the bottom. Rancho, is the hero in this story. He has made his friends to wake up that they should follow his heart and never be afraid of anything. "All izz well", say it when problem come to you. In a short, his friends realized and move on with his interest and the are become successful even not rich but happy.

As usual, I always cry when watching Indian movies. It was really great movie. "All izz well"

A surprise news....

It was a very busy day when I spent my 31st b'day in my working place. "Happy B'day to Asti" was written on the noticeboard and almost all my friends gave me lots of greetings. Unfortunately, plenty of works have made us came back to Wollongong very late at around 6.00 pm on that day. I was very tired when entering my home. Suddenly it was gone when I saw my clean house and welcome smiles of Lita and my beloved hubby Andi. He has cooked my favorite dishes; sate lilit, lawar, sambal sereh and nasi kuning. All of them have been served on the table. Lita also gave me a rose and her own card saying "Happy Birthday to Asti". My happy night completed after 30 minutes latter my best friend family came brought a birthday cake and spent the night together.

Another surprise was coming when I checked my email on that night. "Dear Dr. Palupi, I am pleased to inform you that you have been selected to participate and granted a fellowship by TDR?WHO Geneva for the Advance course of Immunology, vaccinology and biotechnology applied to TB". This email has made my plan change in the very short time. I should leave Andi behind to finish his study without us and return to Jogja soon. My mind think that God has a new plan for me and question come to me: is this the best for us?....

Funding for New Diagnostics of MDR-TB

As repoted in http://www.worldcarecouncil.org/content/finally-good-news-mdr-tb-diagnostics-are-coming by Comrades, there is a new agreement signed between UNITAID, WHO, FIND, GLI and GDF for supporting the new diagnostics for MDR TB.

MDR TB (Multi Drugs Resistance of Tuberculosis), a condition that there are resistance cases to more than one TB drugs that caused TB treatment unsuccessful. This has made the case increased and needed new diagnostic for MDR TB and new drugs as well.

The project called EXPAND TB (Expanding access to new diagnostics for TB) which goal is to accelerate access to diagnosis for patients at risk of MDR TB. These are the partners:

UNITAID, is giving fund $ 26 million and will be expanded to $ 87 million for the project. The 16 targeted countries (2008 - 2011) are: Azerbaijan, Bangladesh, Côte d'Ivoire, the Democratic Republic of the Congo, Ethiopia, Indonesia, Georgia, Kazakhstan, the Kyrgyz Republic, Myanmar, Lesotho, the Republic of Moldova, Tajikistan, Ukraine, Uzbekistan and Viet Nam. The next project (2011-2013) will be included 11 other countries: Cameroon, Djibouti, Haiti, India, Kenya, Peru, Uganda, the United Republic of Tanzania, Senegal, Swaziland and Zambia.

This is a good opportunity for Indonesia as a high burden countries for TB cases in world to strengthen the Laboratory access for new diagnostics. Supplyng the tools and training for laboratory test.