REFERENCE ID: 2018-05278




is a highly prevalent chronic infectious disease caused by Mycobacterium tuberculosis
bacilli (1). Tuberculosis is the
world’s second most common cause of death from infectious disease, after HIV/AIDS
(2). Every year approximately 2
million persons in India develop tuberculosis , accounting for one fourth of
the world’s new Tuberculosis cases (2).
The tuberculosis infection spreads through frequent contact with the infected
person, poor nutrition and living in crowded or unhygienic conditions.

Studies have
documented a close relationship between acid fast stain bacilli in sputum and
hematological abnormalities and revealed that abnormal hematological values are
useful indicators and may aid in diagnosis of severity in TB(7). Anemia is the most common
hematological manifestation during tuberculosis. Active tuberculosis also
produces signs and symptoms that include hematological abnormalities,
hypernatremia and psychological disorders (6).

is a chronic infectious disease, so anemia of inflammation may contribute significantly
(8, 9, and 10). A number of studies
have documented anemia in patients with TB (11,
12). The comprehensive
investigations on hematological changes and abnormalities associated to tuberculosis
are still lacking (13).

Anemia of
chronic disease also called as “anemia of inflammation” is the second most
prevalent after anemia caused by iron deficiency, occurs in patients with acute
or chronic immune activation. Anemia in tuberculosis is most often due to
nutritional deficiency, malabsorption syndromes, failure of iron utilization,
and bone marrow suppression. The most common is normocytic normochromic anemia
of chronic disease. Anemia is more frequently associated with female and oldage.
TB associated anemia is usually mild and resolves with anti TB treatment.



To estimate the level of hemoglobin in the blood of positive tuberculosis
patient above 16 yrs.

To determine the abnormal RBC indices (PCV, MCV, MCH, MCHC) in
tuberculosis patients.

To find out the type of anemia caused in tuberculosis(Normocytic
normochromic  anemia)

To determine platelet count in patients suffering from the disease.

To determine total leucocyte count (TLC) and percentage distribution of
different types of WBCS in a stained Film.




The study
will include 100 tuberculosis patient diagnosed either through sputum
examination or chest X-ray findings. All tuberculosis diagnosed patient falling
in DOTS (Directly observed Treatment, Short Course) will be selected from
outpatient department (OPD) of Dr.D.Y.Patil medical college, hospital &
Research Centre, Pimpri, Pune from May, 2018 to September, 2018. Sputum
positive smears and abnormal chest X-ray are used to diagnose positive
tuberculosis patients in the hospital.


age group of 16 years


16 years



suffering from chronic disease like Diabetes Mellitus, Renal or liver malignancy.

The analysis
of hematological parameters (RBC Count, Hb content, ESR, PCV, MCV, MCH, MCHC, DLC,
TLC and Platelet count) will be done from CCL of hospital. The values will be
measured by automated Cell counter (Sysmex KX- 21) which is based on principle
of Electrical Impudence Counters.

RBC Count: Normal value: Males- 5 to 6.5 million/

3.5 to 5 million/cmm

Hb Content: Normal value : Males- 15-17 gm/100ml
of blood

Females- 13-14.5 gm/100ml of blood

carrying capacity per gram of Hb is 1.34 cc.

Iron content
per gram of Hb is 3.34 mg.

According to
World Health Organization (WHO), the hemoglobin concentration less than 13g/dL
in men and 12 g/dL in women is considered as anemia.

Erythrocyte Sedimentation Rate (ESR):
Normal Range-

 Males: 0-9 mm at the end of 1st
hour (Wintrobe)

               3-5mm at the end of 1st hour

0-20 mm at the end of 1st hour (Wintrobe)

                 4-7 mm at the end of 1st
hour ( Westergren)

Packed cell Volume (PCV): It is the concentration of RBCs. When
PCV is less than normal it indicates normocytic or microcytic anemia.

Normal range

Males     : 40%-47%

Females:  37%-42%

Mean Corpuscular Volume (MCV): It represents average volume of red

Normal range
of MCV: 78-94 cubic microns

Mean Corpuscular Hemoglobin (MCH): It represents average weight of
hemoglobin in one red cell.

Normal range
of MCH: 27-32 picogram

Mean Corpuscular Hemoglobin
Concentration (MCHC):
It represents the amount of hemoglobin per 100 ml of red cells.

Normal range
of MCHC: 32-38%

Total Leucocyte count (TLC): Normal Range: 4000 to 11000 per cmm

Platelet count: Normal range: 1.5 – 4 Lacs/cumm

Differential Leucocyte count (DLC): Distribution of different types of
WBCs in a stained film.


(Polymorphs): 50-70%

Lymphocytes                       : 20-40%

Monocytes                           : 2-8%

Eosinophils                           : 1-4%

Basophils                              : 0-1%


BLOOD SAMPLE COLLECTION: Venous blood samples will be obtained
from patients who are sputum positive and have abnormal chest X-rays. 10 cc of
venous blood will be collected from the patient with aseptic conditions. The
blood will be transferred into a tube containing 0.2 ml of 4% ethylene diamine tetra
acetic acid (EDTA) solution and analyzed in hematology analyzer for evaluation
for different blood parameters. Blood will be also used for erythrocytic
sedimentation rate (ESR) determination. Peripheral blood smear test (PBS) will
be done using abnormal results. In PBS test, blood droplet is spread thinly
into the glass slide and then it is treated with leishman’s stain which fixes
the smear to the slide.  Distilled water
is added to the stain and poured off after few minutes and washed under running
water. Smear is dried and will be examined under oil­-immersion objective of
the microscope.

BACTERIOLOGICAL ANALYSIS: Sputum samples will be collected
from the patients; smears will be stained by Ziehl-Neilsen method (ZN stain)
and seen under the microscope. ZN staining is done by covering fixed smear with
carbol fuschsin, heating gently, decourising with 20% sulphuric acid, counter
staining with methylene blue, wash blot dry and examining under oil immersion

STATISTICAL ANALYSIS: Tables and Graphs will be prepared
by using the collected data. By using different formulas, mathematical
calculations will be performed and results will be obtained.



study of abnormal hematological values will act as useful indicators and will
help in diagnosis of severity in Tuberculosis.

study will demonstrate  a close
relationship between acid fast stained bacilli in sputum and hematological




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